Identification, Genomic Characterization, and Phylogenetic Relationship of the Heat Shock Protein Beta-1 (HSPB1) in Placental Mammals

Identification, Genomic Characterization, and Phylogenetic Relationship of the Heat Shock Protein Beta-1 (HSPB1) in Placental Mammals | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Identification, Genomic Characterization, and Phylogenetic Relationship of the Heat Shock Protein Beta-1 (HSPB1) in Placental Mammals Mehwish Sultana, Shakeela Parveen, Ayesha Khan, Muhammad Abdul Manan, and 5 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6417248/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Heat Shock Protein Beta-1 (HSPB1), a molecular chaperone crucial for celluar response and proteostasis, exhibits evolutionary conservation woth potential lineage-specific adaptations in placental mammals, warranting detailed comparative genomic investigation. The study investigated the characteristics, evolutionary links, motifs, secondary structure, and genetic organization of Heat Shock Protein Beta-1 ( HSPB1) protein across twelve distinct mammals. Significant sequence conservation was identified using multiple sequence alignments (MSA), with over 70% identity in specific areas among the chosen organisms. Physiochemical analysis revealed that all species’ protein sequences exhibited an acidic nature, while instability indices indicated inherent protein instability. The GRAVY analysis referred to hydrophilic properties, while the aliphatic index showed heat stability. Phylogenetic analysis revealed five distinct clades, corresponding to major placental mammals’ groups (e.g. Homo sapiens, Bos Taurus ), which underscores deep evolutionary divergences and conserved stress-response adaptations across lineages. Motif analysis revealed distinctive patterns in several species, and InterProScan results revealed membership in the "Homologous superfamily HSP20_like_Chapserson" family. An examination of the genetic organization indicated differences among organisms in the upstream, downstream, intron, and CDS regions, and the presence of conserved regions suggested their identity and similarity matrices. The current study conducted a computational approach and supporting evidence that HSPB1 is a novel heat shock responsive protein identified in placental mammals The current study findings provide a foundational framework delving into HSPB1 evolutionary and lineage-specific diversification, offering valuable insights into stress adaptation mechanisms and their implications for biomedical or evolutionary studies in mammals. Animal Science Domain Gene structure Genomics HSPB1 Motif Phylogeny Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Figure 6 Introduction In response to environmental stress, all organisms— ranging from archaea and bacteria to plants and animals—produce heat shock proteins. A subgroup of these heat shock proteins is known as small heat shock proteins (sHSPs). HSPB1 also referred toknown as Heat Shock Protein 27, is a member of of this family of sHSPs. The production of these Heat Shock Protein (HSPs) is triggered when cells are exposed to elevated temperatures [ 1 ].The sHSP family is the most prevalent class of molecular chaperones, which are proteins responsible for assisting in the proper folding or unfolding of other proteins and the assembly and disassembly of macromolecular structures,. The functions of molecular chaperones, particularly sHSPs, is critically dependent on their structural properties. sHSPs typically from large oligomeric complexes that resemble spheres or barrels. These Oligomers are macromolecular complexes composed of multiple monomers, usually ranging from 12 to 24 monomers, formed through non-covalent interactions between proteins [ 2 ]. Structural studies, including crystallography, reveal that dimers are the fundamental building blocks that aggregate to form these larger oligomers [ 3 ]. This oligomerization process is essential for the chaperones function of sHSPs, which involves preventing the aggregation of misfolded proteins and assisting in their proper refolding under stress condition [ 4 ]. sHSP are present in virtually all organisms, and their role in cellular protection under stress has led to increasing interest from researchers in recent years To facilitate the study of sHSPs[ 5 ], a dedicated database, known as sHSPdb has been developed. This database serve as a resource to analyze the structure, function and evolving research trends related to sHSPs [ 6 ].Heat Shock Protein 27, also known as HSPB1, is a component of the family of proteins known as small heat shock proteins, or sHSPs. However, stress situations such as heat shock can cause the cell to become exposed, which in turn leads to the folding of proteins and stimulates the production of HSPB1.The production of HSPB1 in the cell results in increased resistance of the cell to the damaging effects of heat shock and oxidative stress. HSPB1 are molecular chaperones, which are proteins that aid in the conformational folding or unfolding as well as the construction or disassembly of other macromolecular structures, and they share an ATP independent holdase activity [ 7 ]. HSPB1 are ATP-independent chaperon; they engage with proteins that have not fully folded, and it is this association that prevents the proteins from aggregating under stressful conditions [ 8 ] and promotes the storage of the proteins in a refolding competent state. In the event that these partially folded or misfolded proteins do not interact with HSPB1, then this leads to irreversible protein aggregation, which can be harmful to cells. In addition to this, HSPB1 is particularly significant since it is associated with diabetic kidney disease as well as viral infection [ 9 , 10 ]. Osteoblasts are very critical cells for the development of bones, and HSPB1 plays a role in the functionality of osteoblasts. Both TNF- and IL-6 are considered to be inflammatory cytokines, and both become active during the process of inflammation. Interleukin (IL)-6 plays an important role in the development of an immune response, the formation of B cells, and the production of neutrophils in the bone marrow. It is also produced when tissue is damaged or infected. The production of tumor necrosis factor alpha (TNF-a), which has a role in the development of resistance to infection and cancer, takes place during acute inflammation. In osteoblast cells such as MC3T3-E1, HSPB1 acts as a regulator for the TNF–stimulated production of IL-6 [ 11 ]. The current study will provide vital information pertaining to the structure, functions, and evolutionary links of the heat shock protein (HSPB1) among mammals. This work aims to systematically identify and characterize HSPB1 orthologs across placental mammals by analyzing their genomic features, evolutionary conservation and lineage-specific adaptations. Additionally, it seeks to elucidate the phylogenetic relationships of HSPB1, tracing its functional divergence and role in stress adaptation mechanisms within eutherian lineages. Materials and Methods Data collection The CDS, protein, and DNA sequences were retrieved from the National Centre for Biotechnology Information (NCBI) https://www.ncbi.nlm.nih.gov/ [ 12 ]. The data for all organisms are shown in Table 1 , together with their respective accession ids, protein ids, and the databases from which they were downloaded. Table 1 Information about data source, organism nucleotide and amino acid sequence length, and accession ID. Species Protein-ID Length Database Accession-ID Database Ochotona princeps XP_004587250.1 233 NCBI XM_004587193.1 NCBI Cavia porcellus XP_003470158.1 200 NCBI XM_003470110.4 NCBI Lynx Canadensis XP_030157330.1 205 NCBI XM_030301470.1 NCBI Bos Taurus NP_001020740.1 204 NCBI NM_001025569.1 NCBI Ovis aries XP_027817273.1 201 NCBI XM_027961472.2 NCBI Camelus ferus XP_032315487.1 201 NCBI XM_032459596.1 NCBI Capra hircus XP_017896392.1 201 NCBI XM_018040903.1 NCBI Pteropus Alecto XP_006918629.1 207 NCBI XM_006918567.3 NCBI Homo sapiens NP_001531.1 205 NCBI NM_001540.5 NCBI Gorilla gorilla XP_004045665.1 205 NCBI XM_004045617.3 NCBI Erinaceus europaeus XP_007518007.1 199 NCBI XM_007517945.2 NCBI Suncus etruscus XP_049643837.1 209 NCBI XM_049787880.1 NCBI Multiple sequence alignment All of the HSPB1- protein sequences were aligned using Multiple Align Show ( https://www.bioinformatics.org/sms/ multi_align.html) [ 13 ], which allowed for the detection and visualization of sequence variants as well as insertions and deletions. Phylogenetic tree The neighbor joining (NJ) approach was used to generate a phylogenetic tree of closely related species using MEGA 11 [ 14 ]. The bootstrap was set to 1000 repeats, and the tree was constructed using this method. ITOL version 4.2.4 was utilized in order to visualize and investigate the findings of phylogeny and multiple sequence alignment. Motif and domain analyses The MEME (Multiple Expectation Maximization) tool [ 15 ] was utilized in order to locate motifs within the protein sequences of each and every organism. The generation of motifs permitted any number of repetitions, and the maximum number of motifs that could be used was 10. In order to locate domains in each and every organism, the InterProScan [ 16 ] was utilized. It was possible to determine which protein families an organism belonged to and, as a result, whether or not it included any functional domains. Gene structure analysis The application of Gene Structure Display Server (GSDS) version 2.0 [ 17 ] was utilized in order to determine the configuration of conserved elements, exons, and introns, as well as their respective localities. Physiochemical properties ProtParam an online tool [ 18 ] that is available on the Expert Protein Analysis System (Expasy) Server which was used to compute physiochemical properties, including the molecular weight (MW), amino acids (AA), theoretical pI, instability index (II), aliphatic index (AI) and grand average of hydropathy (GRAVY). The WOLFPSORT software is used to determine where in the subcellular environment each and every organism's protein resides. The Hydropathy Plot was created with the support of the ProtScale software and then analyzed further. Structural characterizations Using a method for the prediction of secondary structures called GORIV [ 19 ], which is used to calculate the secondary structure parameters (Alpha helix, Beta Bridge, Beta Turn, Extended strand, and Random Coil). The properties of all of an organism's secondary structures are listed in Table 5 . The 3D model of protein was generated using online tool Phyre2, a completely web-based homology modelling tool for protein structures. Results Multiple sequence alignment The Multiple Sequence Alignment (MSA) of each organism's protein was analyzed and displayed using the Multiple Align show. In Fig. 1 , the light red color represents the identical amino acid, while the blue color represents amino acids that are related. Although the color light yellow indicates that there are no similarities between the organisms. More than 70 percent of the sequences are identical along these locations that have been highlighted. Physiochemical analysis The results showed that there were changes in the physiochemical parameters of the organisms (Table 2 ). The species E. europaeus has the lowest number of amino acids (199), while the species O. princeps had the highest number of amino acids (233). Because all of the organisms have theoretical PI values that are lower than 7, it may be deduced that these are acidic in their natural state. All of the organisms' instability indices were higher than 40, indicating that the proteins of all of the selected organisms were unstable. The aliphatic index is a measure that may be used to evaluate the thermal stability of proteins; in this case, its value was greater than 50% for every protein. This suggests that aliphatic side amino acids covered over half of the volume of these proteins. The fact that the GRAVY values were less than 0 demonstrates that the hydrophilic quality of proteins. It was noted that the nucleus and the mitochondria both contain proteins from the organisms that were studied. Additionally, It was observed that the proteins from certain organisms are water-soluble. The hydropathy plot, generated for all of the species and shown in Fig. 2 , demonstrated these organisms tend to exhibi a greater affinity for hydrophobicity. The physiochemical parameters of each organism are provided in Table 2 , including the amino acid composition, extinction coefficient, theoretical pI, instability index, aliphatic index, molecular weight, grand average of hydropathicity (GRAVY) and localization of each protein. Table 2 Physiochemical characteristics of different organisms Species AA length Molecular Weight Theoretical PI Instability index Aliphatic index Gravy Localization Ochotona princeps 233 25476.85 6.60 58.61 77.47 -0.289 Mitochondria Cavia porcellus 200 22284.03 6.12 61.85 68.25 -0.562 Nuclear Lynx Canadensis 205 22720.55 6.23 65.03 67.61 -0.524 Mitochondria Bos Taurus 204 22679.34 5.77 59.53 69.36 -0.597 Mitochondria Ovis aries 201 22334.03 6.22 63.46 70.40 -0.568 Nuclear Camelus ferus 201 22410.17 6.09 65.52 70.85 -0.551 Nuclear Capra hircus 201 22349.00 6.22 62.71 68.46 -0.604 Nuclear Pteropus alecto 207 22853.71 6.32 73.49 66.43 -0.549 Nuclear Homo sapiens 205 22782.52 5.98 62.82 68.54 -0.567 Nuclear Gorilla gorilla 205 22782.52 5.98 62.82 68.54 -0.567 Nuclear Erinaceus europaeus 199 22032.94 6.08 68.37 75.43 -0.438 Nuclear & Mitochondria Suncus etruscus 209 22988.85 6.22 69.20 65.84 -0.492 Nuclear & Mitochondria Phylogenetic analysis Within the evolutionary tree that was constructed with MEGA 11, five distinct groups, or clades, emerged. Figure 3 illustrates quite clearly that members of clade 1, represented by the blue color, S. etruscus and E. europaeus , have descended from the same ancestor and are closely related to one another. In the second group, represented by the color red, H. sapiens and G. gorillas are closely related to one another since they share a common ancestor. C. porcellus , L. canadensis , and O. princeps are the three species that originated from the same ancestor and make up the third clade, which is colored peach. All four of these species, B. taurus , O. aries, C. ferus, and C. hircus , belong to the same clade and are closely related to one another. P. alecto is not closely related to any other organism and is depicted as a single entity in the phylogenetic tree even though it is located in clade 5, which is denoted by the branch color purple. Motif analysis The investigation that was carried out with the assistance of the MEME tool revealed that motifs 1 through 7 were existing in the protein sequences of all species with the exception of E. europaeus . Only the protein sequences of two different species, namely S. etruscus and O. princeps , contained the Motif 8 pattern. Two more species, namely P. alecto and O. princeps , were identified to have a motif 9 in their DNA. Motif 10 was unique to O. princeps , as illustrated in Fig. 4 , and was only identified in that species. Domain analysis According to the results of the InterProScan study, all of the proteins from the species that were selected are members of the same homologous family called "Homologous super family HSP20_like_Chaperson," having the accession number IPR008978. The accession numbers (IPR002068 and IPR037876) for both of the DNA building domains known as "ACD_HSPB1 and A Crystalline" for each species as shown in Table 3 . Table 3 Positions of homologous super families and domains present in the selected organisms Organism Name Homologous Super family HSP20_like_Chaperone IPR008978 Domain 1 ACD_HSPB1 IPR037876 Domain 2 A Crystalline IPR002068 Ochotona princeps 69–189 83–168 75–183 Cavia porcellus 62–187 79–164 71–179 Lynx canadensis 69–191 76–184 84–169 Bos Taurus 67–187 72–180 80–165 Ovis aries 67–187 72–180 80–165 Camelus ferus 65–187 80–165 72–180 Capra hircus 67–187 72–180 80–165 Pteropus alecto 72–193 78–186 86–171 Homo sapiens 69–197 84–169 76–184 Gorilla gorilla 69–197 84–169 76–184 Erinaceus europaeus 69–190 83–168 75–183 Suncus etruscus 69–190 83–168 75–183 Gene structure analysis The analysis of upstream, downstream, intron, and CDS regions was conducted using GSDS for each species. The term upstream and downstream refer to the relative positions of the genetic information within RNA or DNA, respectively. Specifically, downstream refers to the 3' end of the coding strand, while upstream refers to the 5' end of the coding strand. Both B. taurus and O. princeps possess all four components, upstream, downstream intron and CDS regions. Other species, including C. porcellus, L. canadensis, O. aries, C. ferus, C. hircus, P. alecto, H. sapiens, G. gorilla, E. europaeus, and S. etruscus only possess intron and CDS regions of the gene. Percentage identity and similarity of HSPB1 Gene The percentage of amino acids that are identical to one another and the identity matrix of amino acids are crucial factors in predicting evolutionary patterns and distinguishing different species. Tables 4 and 5 present the the identity percentage and the similarity percentage of various species, respectively. A higher identity and similarity percentage between species typically indicates a closer relationship, suggesting that their structure and functions are are more likely to be comparable. In particular, sequences with a similarity percentage of approximately 70% are indicative of shared homology, functional similarity, and highly conserved regions HSPB1 gene. This gene plays a crucial role in cellular stress response and protein maintenance. The pair of species exhibiting a similarity of around 70% in their HSPB1 gene likely share similar functions and evolutionary histories. Table 4 The percentage of amino acids identical across different organisms Ochotona princeps Cavia porcellus Lynx Canadensis Bos taurus Ovis aries Camelus ferus Capra hircus Pteropus alecto Homo sapiens Gorilla gorilla Erinaceus europaeus Suncus etruscus Ochotona princeps 100% 76.56% 81.58% 75.31% 76.98% 76.15% 76.56% 75.73% 76.15% 76.15% 71.54% 75.31% Cavia porcellus 76.56% 100% 92.46% 88.28% 90.79% 92.05% 91.21% 87.44% 88.28% 88.28% 81.58% 86.19% Lynx canadensis 81.58% 92.46% 100% 89.12% 92.05% 91.21% 91.63% 90.37% 90.37% 90.37% 83.68% 89.53% Bos taurus 75.31% 88.28% 89.12% 100% 96.65% 93.3% 96.23% 85.35% 86.19% 86.19% 79.91% 82.84% Ovis aries 76.98% 90.79% 92.05% 96.65% 100% 95.81% 99.58% 87.86% 89.12% 89.12% 82% 85.77% Camelus ferus 76.15% 92.05% 91.21% 93.3% 95.81% 100% 95.39% 89.53% 87.86% 87.86% 82.42% 85.35% Capra hircus 76.56% 91.21% 91.63% 96.23% 99.58% 95.39% 100% 87.44% 89.53% 89.53% 81.58% 86.19% Pteropus alecto 75.73% 87.44% 90.37% 85.35% 87.86% 89.53% 87.44% 100% 89.12% 89.12% 83.68% 89.12% Homo sapiens 76.15% 88.28% 90.37% 86.19% 89.12% 87.86% 89.53% 89.12% 100% 100% 82.42% 88.7% Gorilla gorilla 76.15% 88.28% 90.37% 86.19% 89.12% 87.86% 89.53% 89.12% 100% 100% 82.42% 88.7% Erinaceus europaeus 71.54% 81.58% 83.68% 79.91% 82% 82.42% 81.58% 83.68% 82.42% 82.42% 100% 82.84% Suncus etruscus 75.31% 86.19% 89.53% 82.84% 85.77% 85.35% 86.19% 89.12% 88.7% 88.7% 82.84% 100% Table 5 The Percentage of amino acids similar across different organisms Ochotona princeps Cavia porcellus Lynx canadensis Bos taurus Ovis aries Camelus ferus Capra hircus Pteropus alecto Homo sapiens Gorilla gorilla Erinaceus europaeus Suncus etruscus Ochotona princeps 100% 75.73% 80.33% 74.47% 75.73% 75.73% 75.31% 76.56% 75.73% 75.73% 71.12% 76.56% Cavia porcellus 75.73% 100% 78.66% 74.89% 76.15% 78.24% 76.56% 75.73% 75.73% 75.73% 69.45% 75.31% Lynx canadensis 80.33% 78.66% 100% 76.15% 77.82% 77.82% 77.4% 78.24% 77.4% 77.4% 71.12% 77.82% Bos taurus 74.47% 74.89% 76.15% 100% 82% 79.49% 81.58% 74.89% 74.47% 74.47% 68.2% 73.22% Ovis aries 75.73% 76.15% 77.82% 82% 100% 80.75% 83.68% 76.15% 76.15% 76.15% 69.03% 74.89% Camelus ferus 75.73% 78.24% 77.82% 79.49% 80.75% 100% 80.33% 76.98% 75.73% 75.73% 69.45% 75.31% Capra hircus 75.31% 76.56% 77.4% 81.58% 83.68% 80.33% 100% 75.73% 76.56% 76.56% 68.61% 75.31% Pteropus alecto 76.56% 75.73% 78.24% 74.89% 76.15% 76.98% 75.73% 100% 78.24% 78.24% 72.38% 78.66% Homo sapiens 75.73% 75.73% 77.4% 74.47% 76.15% 75.73% 76.56% 78.24% 100% 85.77% 70.71% 78.66% Gorilla gorilla 75.73% 75.73% 77.4% 74.47% 76.15% 75.73% 76.56% 78.24% 85.77% 100% 70.71% 78.66% Erinaceus europaeus 71.12% 69.45% 71.12% 68.2% 69.03% 69.45% 68.61% 72.38% 70.71% 70.71% 100% 73.22% Suncus etruscus 76.56% 75.31% 77.82% 73.22% 74.89% 75.31% 75.31% 78.66% 78.66% 78.66% 73.22% 100% Structural analysis GORIV tool was used to make the prediction about the secondary structure of the HSPB1 gene for the 12 organisms that were chosen. According to Table 6 , the HSPB1 gene has an alpha helix, a random coil, and extended strands as part of its secondary structure. The HSPB1 gene has an alpha helix, a random coil, and extended strands as part of its secondary structure. The L. canadensis alpha helix takes up a total of 26.83 percent of the structure. The portion of the structure that is occupied by the alpha helix, the least by H. sapiens and G. gorilla is 13.66 percent. In addition, the extended strand for the B. taurus was the longest at 17.75%, while the extended strand for the C. porcellus was the shortest at 11.00%. On the other hand, random coils make up 73.17% of the structure of H. sapiens and G. gorilla , which is the largest percentage of random coils among other creatures. For HSPB1 protein sequence, a 12, 3D structure templates were created, and all of the models were successfully constructed from Phyre2 (Fig. 6 ). The 3D structure of HSPB1 protein revealed details about their folding, stability, and domain interactions. Table 6 All the criteria and percentages for the secondary structure of the organisms Organism Name Alpha helix Beta Bridge Beta Turn Extended strand Random Coil Ochotona princeps 26.61% 0.00% 0.00% 14.16% 59.23% Cavia porcellus 21.00% 0.00% 0.00% 11.00% 68.00% Lynx Canadensis 26.83% 0.00% 0.00% 12.20% 60.98% Bos Taurus 21.08% 0.00% 0.00% 17.75% 61.27% Ovis aries 20.90% 0.00% 0.00% 14.93% 64.18% Camelus ferus 19.90% 0.00% 0.00% 16.40% 63.68% Capra hircus 20.90% 0.00% 0.00% 12.94% 66.17% Pteropus Alecto 18.36% 0.00% 0.00% 13.53% 68.12% Homo sapiens 13.66% 0.00% 0.00% 13.17% 73.17% Gorilla gorilla 13.66% 0.00% 0.00% 13.17 73.17% Erinaceus europaeus 16.58% 0.00% 0.00% 17.59% 65.83% Suncus etruscus 18.66% 0.00% 0.00% 12.44% 68.90% Discussion Heat shock protein 27, also known as HSPB1, is a member of the family of proteins known as small heat shock proteins, or sHSPs. This protein contributes to the development of a stable state during times of high stress, such as when the body is subjected to heat shock or oxidative damage [20]. The findings of this study were derived from a computational analysis of the HSPB1 protein found in a variety of species. During this computational analysis, a number of analyses were carried out in order to identify Domains, multiple sequence alignment, physiochemical properties, motifs, quaternary structure, secondary structure parameters, protein interaction with its functional proteins in a variety of organisms, similarity and identity matrix, and the Position and composition of conserved elements, introns, exons in protein. A gene family's genetic characterization can reveal details about the genes' origins, duplication events, and diversification throughout their evolutionary history. The conserved regions, which are crucial for protein structure and functional studies, can be found using multiple sequence alignment (MSA) [21]. In nucleus majority of the proteins were found, while the mitochondria only contained a small percentage. Except for B. taurus and O. princeps , all creatures had just CDS parts, downstream and upstream in their protein sequences. These two species have CDS parts, Introns, downstream and upstream in their protein sequences. When referring to the relative placement of the genetic code in DNA or RNA, the terms "downstream" and "upstream" are both used. Upstream is towards the 5' end of the coding strand of the protein that is now being analyzed, while downstream is towards the 3' end of the coding strand. Both ACD_HSPB1 IPR037876 and A Crystalline IPR002068 were identified as the domains, present in each and every organism. The homologous superfamily found in all organisms was designated as IPR008978 Hsp20. The motif from 1 to 7 was found in the protein sequences of all organisms, with the exception of E. europaeus . Only the protein sequences of two different organisms, namely S. etruscus and O. princeps , contained the Motif 8 pattern. Two more organisms, namely P. alecto and O. princeps , were found to have a motif 9 in their DNA. While Motif 10 was unique to O. princeps , it was detected in both species. Furthermore, the varied patterns of the introns and downstream/upstream untranslated regions (UTRs) in the gene structure suggest that it may be due to the inclusion and removal of retroposons [22]. A prior study [23] was also responsible for collecting amino acids, determining % similarity and identity, phylogenetic relationship, determining physiochemical qualities, and identifying motifs. The phylogenetic tree revealed the presence of five distinct clades. In clade 1(represented by blue color), the species S. etruscus and E. europaeus share common ancestor, indicating their close evolutionary relationship. In the second clade (marked in red), H. sapiens and G. gorillas are closely related, as evidenced by to their shared common ancestor. The third clade, colored peach, consists of C. porcellus, L. canadensis, and O. princeps, which all originated from a common ancestor, highlighting their evolutionary proximity. The fourth clade includes B. taurus , O. aries, C. ferus , and C. hircus , which are all closely related, stemming from a shared lineage. Lastly, P. alecto is depicted as a distinct entity in clade 5 (marked in purple), isolated from other species, despite being located within this clade. The phylogenetic analysis also revealed a high degree of sequence homology among the species, reinforcing their evolutionary relationships as observed in other studies [24]. Furthermore, assessing the physiochemical characteristics of the gene encoding the gene is necessary to comprehend the protein structure and function of the gene family [25]. According to the findings, every organism has a greater tendency towards hydrophilicity. The aliphatic index is a measure that may be used to evaluate the thermal stability of proteins; in this case, its value was greater than 50% for every protein. This suggests that aliphatic side amino acids covered over half of the volume of these proteins. In addition, GRAVY values that are lower than zero indicate that the hydrophilicity of these proteins is greater than that of their surroundings. The fact that all of the organisms have theoretical PI values that are lower than 7 demonstrates that they are all acidic in their natural state. Because the instability index for every creature was greater than 40, it may be deduced that every protein in every organism, with one exception, is unstable. Similarly, since GRAVY indicates a protein's hydrophilicity (GRAVY 0), assessing it would aid in a better understanding of its tertiary structure or shape [26]. Similar genetic characterization was done in previous studies including the assembly of amino acids and nucleotides, the computation of similarity and identity matrices, the assessment of physiochemical properties, the analysis of evolutionary relationships, the characterization of structural features, and the generation of a hydrophobicity plot [27]. Hence in recent years, determining and quantifying the evolutionary processes that contributed to genetic variety has emerged as an exciting area of research. The secondry structure of the HSPB1 gene in 12 species was predicted using GORIV tool, indicating the presence of alpha helices, random coils and extended strands. L. canadensis showed the highest alpha helix content, while H. sapiens and G. gorillas had the largest proportion of random coils. The three-dimensional models generated for the HSPB1 protein provided insights into its folding. Stability and domain interactions. These findings align with the previous studies on protein structure-function relationship and emphasize the evolutionary conservation of the HSPB1 gene across the species [28],[29]. Conclusion This study provides comprehensive insights into the evolutionary history, functional motifs, physiochemical characteristics and structural traits of the HSPB1 gene across various species. The results revealed the vital role of HSPB1 proteins in cellular processes, emphasizinf their acidic nature and high sequence conservation. Phgylogenetic analysis highlighted the evolutionary relationship among species, depicting their shared ancestry, The GRAVY values pointed to the hydrophilic nature of these proteins while the aliphatic index indicated their heat stability, further emphasizing their functional significance. The study also unveiled conserved motifs and homologous superfamily memberships which underscore the functional conservation of HSPB1 gene across the species. Variations in genetic organization, like the presence of distinct upstream and downstream regions in certain species reflect the complexity of gene regulation. Furthermore, the presence of conserved regions within the supports the notion of functional similarity across the species. The identity and similarity matrices confirmed that the species with higher similarity percentages in the HSPB1 gene likely shre common functions and evolutionary backgrounds. The analysis of secondary structure, including the distribution of alpha helices, random coils and extended strands, provided valuable insights on the protein folding, stability and domain interactions, the three-dimensional structure of HSPB1 protein offered further insights into its functional architecture. This work identifies HSPB1 as functionally significant and underscores the need for additional research to fully comprehend the function and mechanism of HSPs. Declarations Conflict of Interest: Authors have declared no conflict of interest. Acknowledgements: None Data Availability Statement: All the data is included in the manuscript. Authors’ Contribution: M.S wrote original draft, S.P conceived main idea, A.K and M.A.M performed analysis, S.J and M. H wrote methodology, S.S and M.T prepared tables, M.F.K revised the manuscript and project administration. References Mymrikov, E.V., A.S. Seit-Nebi, and N.B. Gusev, Large Potentials of Small Heat Shock Proteins. Physiological Reviews, 2011. 91 (4): p. 1123-1159. Russo Krauss, I., et al., An Overview of Biological Macromolecule Crystallization. International Journal of Molecular Sciences, 2013. 14 (6): p. 11643-11691. Salahuddin, P., et al., Structure of amyloid oligomers and their mechanisms of toxicities: Targeting amyloid oligomers using novel therapeutic approaches. European Journal of Medicinal Chemistry, 2016. 114 : p. 41-58. Eyles, S.J. and L.M. Gierasch, Nature’s molecular sponges: Small heat shock proteins grow into their chaperone roles. Proceedings of the National Academy of Sciences, 2010. 107 (7): p. 2727-2728. Jaspard, E. and G. Hunault, sHSPdb: a database for the analysis of small Heat Shock Proteins. BMC Plant Biology, 2016. 16 (1): p. 135. Feng, P., et al., Classifying the superfamily of small heat shock proteins by using g-gap dipeptide compositions. International Journal of Biological Macromolecules, 2021. 167 : p. 1575-1578. Acunzo, J., M. Katsogiannou, and P. Rocchi, Small heat shock proteins HSP27 (HspB1), αB-crystallin (HspB5) and HSP22 (HspB8) as regulators of cell death. The International Journal of Biochemistry & Cell Biology, 2012. 44 (10): p. 1622-1631. Bakthisaran, R., R. Tangirala, and C.M. Rao, Small heat shock proteins: Role in cellular functions and pathology. Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics, 2015. 1854 (4): p. 291-319. Borgo, C., et al., Protein kinase CK2: a potential therapeutic target for diverse human diseases. Signal Transduction and Targeted Therapy, 2021. 6 (1): p. 183. Bolhassani, A. and E. Agi, Heat shock proteins in infection. Clinica Chimica Acta, 2019. 498 : p. 90-100. Mebarek, S., et al., Phospholipases of Mineralization Competent Cells and Matrix Vesicles: Roles in Physiological and Pathological Mineralizations. International Journal of Molecular Sciences, 2013. 14 (3): p. 5036-5129. Coordinators, N.R., Database resources of the National Center for Biotechnology Information. Nucleic Acids Research, 2015. 44 (D1): p. D7-D19. McDonald, E.T., et al., Sequence, Structure, and Dynamic Determinants of Hsp27 (HspB1) Equilibrium Dissociation Are Encoded by the N-Terminal Domain. Biochemistry, 2012. 51 (6): p. 1257-1268. Baharum, S. and A.w.A. Nurdalila, Phylogenetic Relationships of Epinephelus fuscoguttatus and Epinephelus hexagonatus Inferred from Mitochondrial Cytochrome b Gene Sequences using Bioinformatic Tools. International Journal of Bioscience, Biochemistry and Bioinformatics, 2011. 1 (1): p. 47. Bailey, T.L., et al., The value of position-specific priors in motif discovery using MEME. BMC Bioinformatics, 2010. 11 (1): p. 179. Blum, M., et al., The InterPro protein families and domains database: 20 years on. Nucleic Acids Research, 2020. 49 (D1): p. D344-D354. Iqbal Qureshi, A.M., et al., Insilco identification and characterization of superoxide dismutase gene family in Brassica rapa. Saudi Journal of Biological Sciences, 2021. 28 (10): p. 5526-5537. Sahay, A., A. Piprodhe, and M. Pise, In silico analysis and homology modeling of strictosidine synthase involved in alkaloid biosynthesis in catharanthus roseus. Journal of Genetic Engineering and Biotechnology, 2020. 18 (1): p. 44. CoSec: a hub of online tools for comparing secondary structure elements. International Journal of Bioinformatics Research and Applications, 2023. 19 (1): p. 56-69. Kurashova, N.A., I.M. Madaeva, and L.I. Kolesnikova, Expression of HSP70 Heat-Shock Proteins under Oxidative Stress. Advances in Gerontology, 2020. 10 (1): p. 20-25. Sultana, M., et al., In silico molecular characterization of TGF-β gene family in Bufo bufo: genome-wide analysis. Journal of Biomolecular Structure and Dynamics: p. 1-15. Su, J., et al., Comparative evolutionary and molecular genetics based study of Buffalo lysozyme gene family to elucidate their antibacterial function. International Journal of Biological Macromolecules, 2023. 234 : p. 123646. Akbari Rokn Abadi, S., et al., An accurate alignment-free protein sequence comparator based on physicochemical properties of amino acids. Scientific Reports, 2022. 12 (1): p. 11158. Hassan, F.-u., et al., Genome-wide identification and evolutionary analysis of the FGF gene family in buffalo. Journal of Biomolecular Structure and Dynamics, 2024. 42 (19): p. 10225-10236. Lee, D., O. Redfern, and C. Orengo, Predicting protein function from sequence and structure. Nature Reviews Molecular Cell Biology, 2007. 8 (12): p. 995-1005. Amrhein, S., et al., Molecular Dynamics Simulations Approach for the Characterization of Peptides with Respect to Hydrophobicity. The Journal of Physical Chemistry B, 2014. 118 (7): p. 1707-1714. Das, J.K., et al., Mapping sequence to feature vector using numerical representation of codons targeted to amino acids for alignment-free sequence analysis. Gene, 2021. 766 : p. 145096. M. F. Khan et al. , “Evolution and comparative genomics of the transforming growth factor-β-related proteins in Nile tilapia,” Mol Biotechnol , pp. 1–15, 2024. S. Parveen, M. F. Khan, M. Sultana, S. ur Rehman, and L. Shafique, “Molecular characterization of doublesex and Mab-3 (DMRT) gene family in Ctenopharyngodon idella (grass carp),” J Appl Genet , pp. 1–12, 2024. Additional Declarations The authors declare no competing interests. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6417248","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":440986884,"identity":"03e3afe9-76b1-4227-8ffd-ac30e41ebf60","order_by":0,"name":"Mehwish Sultana","email":"","orcid":"","institution":"Department of Zoology, The Government Sadiq College Women University, 63100, Bahawalpur, Punjab, Pakistan.","correspondingAuthor":false,"prefix":"","firstName":"Mehwish","middleName":"","lastName":"Sultana","suffix":""},{"id":440986885,"identity":"eb1548ae-ce12-433a-a541-e19f53c277d5","order_by":1,"name":"Shakeela Parveen","email":"","orcid":"","institution":"Department of Zoology, The Government Sadiq College Women University, 63100, Bahawalpur, Punjab, Pakistan.","correspondingAuthor":false,"prefix":"","firstName":"Shakeela","middleName":"","lastName":"Parveen","suffix":""},{"id":440986886,"identity":"7f01d79e-e72a-4ff1-8772-1888840815dd","order_by":2,"name":"Ayesha Khan","email":"","orcid":"","institution":"Department of Zoology, Punjab University, Lahore, Pakistan.","correspondingAuthor":false,"prefix":"","firstName":"Ayesha","middleName":"","lastName":"Khan","suffix":""},{"id":440986887,"identity":"657d3b4f-a383-417b-ba18-24e04744a188","order_by":3,"name":"Muhammad Abdul Manan","email":"","orcid":"","institution":"Department of Zoology, Punjab University, Lahore, Pakistan.","correspondingAuthor":false,"prefix":"","firstName":"Muhammad","middleName":"Abdul","lastName":"Manan","suffix":""},{"id":440986888,"identity":"61295189-3f38-45b3-9e54-2ed4d494e548","order_by":4,"name":"Sarwar Jahan","email":"","orcid":"","institution":"Higher Education Department, Khyber Pakhtunkhwa, Peshawar, Pakistan.","correspondingAuthor":false,"prefix":"","firstName":"Sarwar","middleName":"","lastName":"Jahan","suffix":""},{"id":440986889,"identity":"2629e645-9d41-4b97-b696-c90301110744","order_by":5,"name":"Muhammad Hussain","email":"","orcid":"","institution":"Department of Veterinary and Animal Sciences, University of Veterinary and Animal Sciences, Lahore,","correspondingAuthor":false,"prefix":"","firstName":"Muhammad","middleName":"","lastName":"Hussain","suffix":""},{"id":440986890,"identity":"6cac5e50-51f2-4656-9ce2-538496582453","order_by":6,"name":"Saba Saeed","email":"","orcid":"","institution":"Department of Zoology, The Government Sadiq College Women University, 63100, Bahawalpur, Punjab, Pakistan.","correspondingAuthor":false,"prefix":"","firstName":"Saba","middleName":"","lastName":"Saeed","suffix":""},{"id":440986891,"identity":"9311a3d6-4ede-42f8-8745-baeb29ce62ab","order_by":7,"name":"Muhammad Tayyab","email":"","orcid":"","institution":"Department of Zoology, Government College University Faisalabad, Punjab, Pakistan.","correspondingAuthor":false,"prefix":"","firstName":"Muhammad","middleName":"","lastName":"Tayyab","suffix":""},{"id":440986892,"identity":"5c28b06f-9709-41d7-bfcd-5789f9bca3bf","order_by":8,"name":"Muhammad Farhan Khan","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA/0lEQVRIiWNgGAWjYBAC9gYGNiCVkMDAwAOkK4CYmbkBrxaeAyhazoC0MJKihbENJEZIC/vZZw9+7kjL45/de/g177zaaP52oJYfFdtwa+FJNzfsPZNTLHHnXJo177bjuTMOMzYw9py5jVOLPUMamwRvW0Viw40cM2PebcdyG4BamBnbcGvh4X/GJvkXqGU+WMucY7nzCWqRSGOT5m3LSdxwI8f4MW9DTe4GwlqesUnLtqUlbryRl8Y459iB3I1ALQfx+YWHP41N8m1bcuK8G7mHP7ypqcudd/7wwQc/KnBrQQZsEgwMh8GsA0SpBwLmDwwMdcQqHgWjYBSMghEEALaOXOlK17nGAAAAAElFTkSuQmCC","orcid":"","institution":"Department of Chemistry, Gomal University, Dera Ismail Khan, Pakistan.","correspondingAuthor":true,"prefix":"","firstName":"Muhammad","middleName":"Farhan","lastName":"Khan","suffix":""}],"badges":[],"createdAt":"2025-04-10 06:53:59","currentVersionCode":1,"declarations":{"humanSubjects":false,"vertebrateSubjects":true,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":true},"doi":"10.21203/rs.3.rs-6417248/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6417248/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":80361034,"identity":"02f02efc-3ec2-48e1-98f7-845abe00ed55","added_by":"auto","created_at":"2025-04-11 03:58:14","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":351534,"visible":true,"origin":"","legend":"\u003cp\u003eThe multiple sequence alignment of the twelve proteins that was obtained by using Multiple align is displayed here. The protein sequences in the red dark regions share an identity of greater than 70 percent.\u003c/p\u003e","description":"","filename":"1.png","url":"https://assets-eu.researchsquare.com/files/rs-6417248/v1/05cb8e025fbf0743e808070e.png"},{"id":80361033,"identity":"ef0ad639-9fcd-47f0-94b2-39335cc9983d","added_by":"auto","created_at":"2025-04-11 03:58:14","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":171868,"visible":true,"origin":"","legend":"\u003cp\u003eThe hydrophobicity plot for these four creatures illustrates that, in their natural states, they are all more inclined to be hydrophilic.\u003c/p\u003e","description":"","filename":"2.png","url":"https://assets-eu.researchsquare.com/files/rs-6417248/v1/2fca3e8eb75a5439eaeaaf1c.png"},{"id":80361036,"identity":"75390c43-d7ea-4e48-a05d-526e8f5c950b","added_by":"auto","created_at":"2025-04-11 03:58:14","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":452548,"visible":true,"origin":"","legend":"\u003cp\u003eThe phylogenetic tree was constructed by using the maximum likelihood approach in conjunction with the bootstrap test (1000 replications) in MEGA 11 and ITOL.\u003c/p\u003e","description":"","filename":"3.png","url":"https://assets-eu.researchsquare.com/files/rs-6417248/v1/ac156c041de3f91e91196edf.png"},{"id":80361029,"identity":"9b37f25f-ae07-4e49-899d-a7a87b5e231e","added_by":"auto","created_at":"2025-04-11 03:58:14","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":147800,"visible":true,"origin":"","legend":"\u003cp\u003eThe motifs found in the protein sequences of 12 distinct organisms, with each motif represented by a unique color.\u003c/p\u003e","description":"","filename":"4.png","url":"https://assets-eu.researchsquare.com/files/rs-6417248/v1/404744efdedfc5a5c7467baf.png"},{"id":80362006,"identity":"c4d1f4ac-7bdf-4e43-9e41-76a40d755f52","added_by":"auto","created_at":"2025-04-11 04:14:14","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":63408,"visible":true,"origin":"","legend":"\u003cp\u003eA Graphical representation of GSDS indicating proteins that are upstream, downstream, in the intron, and in the CDS portion\u003c/p\u003e","description":"","filename":"5.png","url":"https://assets-eu.researchsquare.com/files/rs-6417248/v1/3d214399777b8dce599e14ad.png"},{"id":80362007,"identity":"2ddae6c5-43a0-44fe-a4ce-cd290b735587","added_by":"auto","created_at":"2025-04-11 04:14:14","extension":"png","order_by":6,"title":"Figure 6","display":"","copyAsset":false,"role":"figure","size":247568,"visible":true,"origin":"","legend":"\u003cp\u003eProtein tertiary structures predicted by Phyre2\u003c/p\u003e","description":"","filename":"6.png","url":"https://assets-eu.researchsquare.com/files/rs-6417248/v1/c910ea26fb2e00ebae0dfc72.png"},{"id":80362298,"identity":"13e587ce-2739-45d6-b2e6-6861a53d359e","added_by":"auto","created_at":"2025-04-11 04:22:16","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2949384,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6417248/v1/4894ae67-43b6-4615-92f1-8b8aa6805ce2.pdf"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"\u003cp\u003e\u003cstrong\u003eIdentification, Genomic Characterization, and Phylogenetic Relationship of the Heat Shock Protein Beta-1 (HSPB1) in Placental Mammals\u003c/strong\u003e\u003c/p\u003e","fulltext":[{"header":"Introduction","content":"\u003cp\u003eIn response to environmental stress, all organisms\u0026mdash; ranging from archaea and bacteria to plants and animals\u0026mdash;produce heat shock proteins. A subgroup of these heat shock proteins is known as small heat shock proteins (sHSPs). HSPB1 also referred toknown as Heat Shock Protein 27, is a member of of this family of sHSPs. The production of these Heat Shock Protein (HSPs) is triggered when cells are exposed to elevated temperatures [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e].The sHSP family is the most prevalent class of molecular chaperones, which are proteins responsible for assisting in the proper folding or unfolding of other proteins and the assembly and disassembly of macromolecular structures,. The functions of molecular chaperones, particularly sHSPs, is critically dependent on their structural properties. sHSPs typically from large oligomeric complexes that resemble spheres or barrels. These Oligomers are macromolecular complexes composed of multiple monomers, usually ranging from 12 to 24 monomers, formed through non-covalent interactions between proteins [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Structural studies, including crystallography, reveal that dimers are the fundamental building blocks that aggregate to form these larger oligomers [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. This oligomerization process is essential for the chaperones function of sHSPs, which involves preventing the aggregation of misfolded proteins and assisting in their proper refolding under stress condition [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e]. sHSP are present in virtually all organisms, and their role in cellular protection under stress has led to increasing interest from researchers in recent years To facilitate the study of sHSPs[\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e], a dedicated database, known as sHSPdb has been developed. This database serve as a resource to analyze the structure, function and evolving research trends related to sHSPs [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e].Heat Shock Protein 27, also known as HSPB1, is a component of the family of proteins known as small heat shock proteins, or sHSPs. However, stress situations such as heat shock can cause the cell to become exposed, which in turn leads to the folding of proteins and stimulates the production of HSPB1.The production of HSPB1 in the cell results in increased resistance of the cell to the damaging effects of heat shock and oxidative stress. HSPB1 are molecular chaperones, which are proteins that aid in the conformational folding or unfolding as well as the construction or disassembly of other macromolecular structures, and they share an ATP independent holdase activity [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. HSPB1 are ATP-independent chaperon; they engage with proteins that have not fully folded, and it is this association that prevents the proteins from aggregating under stressful conditions [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e] and promotes the storage of the proteins in a refolding competent state. In the event that these partially folded or misfolded proteins do not interact with HSPB1, then this leads to irreversible protein aggregation, which can be harmful to cells.\u003c/p\u003e \u003cp\u003eIn addition to this, HSPB1 is particularly significant since it is associated with diabetic kidney disease as well as viral infection [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e, \u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. Osteoblasts are very critical cells for the development of bones, and HSPB1 plays a role in the functionality of osteoblasts. Both TNF- and IL-6 are considered to be inflammatory cytokines, and both become active during the process of inflammation. Interleukin (IL)-6 plays an important role in the development of an immune response, the formation of B cells, and the production of neutrophils in the bone marrow. It is also produced when tissue is damaged or infected. The production of tumor necrosis factor alpha (TNF-a), which has a role in the development of resistance to infection and cancer, takes place during acute inflammation. In osteoblast cells such as MC3T3-E1, HSPB1 acts as a regulator for the TNF\u0026ndash;stimulated production of IL-6 [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. The current study will provide vital information pertaining to the structure, functions, and evolutionary links of the heat shock protein (HSPB1) among mammals.\u003c/p\u003e \u003cp\u003eThis work aims to systematically identify and characterize HSPB1 orthologs across placental mammals by analyzing their genomic features, evolutionary conservation and lineage-specific adaptations. Additionally, it seeks to elucidate the phylogenetic relationships of HSPB1, tracing its functional divergence and role in stress adaptation mechanisms within eutherian lineages.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eData collection\u003c/h2\u003e \u003cp\u003eThe CDS, protein, and DNA sequences were retrieved from the National Centre for Biotechnology Information (NCBI) \u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.ncbi.nlm.nih.gov/\u003c/span\u003e\u003cspan address=\"https://www.ncbi.nlm.nih.gov/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. The data for all organisms are shown in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, together with their respective accession ids, protein ids, and the databases from which they were downloaded.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eInformation about data source, organism nucleotide and amino acid sequence length, and accession ID.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSpecies\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eProtein-ID\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eLength\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eDatabase\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eAccession-ID\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eDatabase\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eOchotona princeps\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eXP_004587250.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e233\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNCBI\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eXM_004587193.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNCBI\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eCavia porcellus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eXP_003470158.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNCBI\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eXM_003470110.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNCBI\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eLynx Canadensis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eXP_030157330.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e205\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNCBI\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eXM_030301470.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNCBI\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eBos Taurus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNP_001020740.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e204\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNCBI\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eNM_001025569.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNCBI\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eOvis aries\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eXP_027817273.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e201\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNCBI\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eXM_027961472.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNCBI\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eCamelus ferus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eXP_032315487.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e201\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNCBI\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eXM_032459596.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNCBI\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eCapra hircus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eXP_017896392.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e201\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNCBI\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eXM_018040903.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNCBI\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePteropus Alecto\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eXP_006918629.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e207\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNCBI\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eXM_006918567.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNCBI\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eHomo sapiens\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eNP_001531.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e205\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNCBI\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eNM_001540.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNCBI\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eGorilla gorilla\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eXP_004045665.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e205\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNCBI\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eXM_004045617.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNCBI\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eErinaceus europaeus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eXP_007518007.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e199\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNCBI\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eXM_007517945.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNCBI\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eSuncus etruscus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eXP_049643837.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e209\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNCBI\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eXM_049787880.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eNCBI\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eMultiple sequence alignment\u003c/h3\u003e\n\u003cp\u003eAll of the HSPB1- protein sequences were aligned using Multiple Align Show (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttps://www.bioinformatics.org/sms/\u003c/span\u003e\u003cspan address=\"https://www.bioinformatics.org/sms/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e multi_align.html) [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e], which allowed for the detection and visualization of sequence variants as well as insertions and deletions.\u003c/p\u003e\n\u003ch3\u003ePhylogenetic tree\u003c/h3\u003e\n\u003cp\u003eThe neighbor joining (NJ) approach was used to generate a phylogenetic tree of closely related species using MEGA 11 [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e]. The bootstrap was set to 1000 repeats, and the tree was constructed using this method. ITOL version 4.2.4 was utilized in order to visualize and investigate the findings of phylogeny and multiple sequence alignment.\u003c/p\u003e\n\u003ch3\u003eMotif and domain analyses\u003c/h3\u003e\n\u003cp\u003eThe MEME (Multiple Expectation Maximization) tool [\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e] was utilized in order to locate motifs within the protein sequences of each and every organism. The generation of motifs permitted any number of repetitions, and the maximum number of motifs that could be used was 10. In order to locate domains in each and every organism, the InterProScan [\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e] was utilized. It was possible to determine which protein families an organism belonged to and, as a result, whether or not it included any functional domains.\u003c/p\u003e\n\u003ch3\u003eGene structure analysis\u003c/h3\u003e\n\u003cp\u003eThe application of Gene Structure Display Server (GSDS) version 2.0 [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e] was utilized in order to determine the configuration of conserved elements, exons, and introns, as well as their respective localities.\u003c/p\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003ePhysiochemical properties\u003c/h2\u003e \u003cp\u003eProtParam an online tool [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e] that is available on the Expert Protein Analysis System (Expasy) Server which was used to compute physiochemical properties, including the molecular weight (MW), amino acids (AA), theoretical pI, instability index (II), aliphatic index (AI) and grand average of hydropathy (GRAVY). The WOLFPSORT software is used to determine where in the subcellular environment each and every organism's protein resides. The Hydropathy Plot was created with the support of the ProtScale software and then analyzed further.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eStructural characterizations\u003c/h3\u003e\n\u003cp\u003eUsing a method for the prediction of secondary structures called GORIV [\u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e], which is used to calculate the secondary structure parameters (Alpha helix, Beta Bridge, Beta Turn, Extended strand, and Random Coil). The properties of all of an organism's secondary structures are listed in Table\u0026nbsp;\u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e. The 3D model of protein was generated using online tool Phyre2, a completely web-based homology modelling tool for protein structures.\u003c/p\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec11\" class=\"Section2\"\u003e \u003ch2\u003eMultiple sequence alignment\u003c/h2\u003e \u003cp\u003eThe Multiple Sequence Alignment (MSA) of each organism's protein was analyzed and displayed using the Multiple Align show. In Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, the light red color represents the identical amino acid, while the blue color represents amino acids that are related. Although the color light yellow indicates that there are no similarities between the organisms. More than 70 percent of the sequences are identical along these locations that have been highlighted.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec12\" class=\"Section2\"\u003e \u003ch2\u003ePhysiochemical analysis\u003c/h2\u003e \u003cp\u003eThe results showed that there were changes in the physiochemical parameters of the organisms (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). The species \u003cem\u003eE. europaeus\u003c/em\u003e has the lowest number of amino acids (199), while the species \u003cem\u003eO. princeps\u003c/em\u003e had the highest number of amino acids (233). Because all of the organisms have theoretical PI values that are lower than 7, it may be deduced that these are acidic in their natural state. All of the organisms' instability indices were higher than 40, indicating that the proteins of all of the selected organisms were unstable. The aliphatic index is a measure that may be used to evaluate the thermal stability of proteins; in this case, its value was greater than 50% for every protein. This suggests that aliphatic side amino acids covered over half of the volume of these proteins. The fact that the GRAVY values were less than 0 demonstrates that the hydrophilic quality of proteins. It was noted that the nucleus and the mitochondria both contain proteins from the organisms that were studied. Additionally, It was observed that the proteins from certain organisms are water-soluble. The hydropathy plot, generated for all of the species and shown in Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, demonstrated these organisms tend to exhibi a greater affinity for hydrophobicity. The physiochemical parameters of each organism are provided in Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e, including the amino acid composition, extinction coefficient, theoretical pI, instability index, aliphatic index, molecular weight, grand average of hydropathicity (GRAVY) and localization of each protein.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePhysiochemical characteristics of different organisms\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"8\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSpecies\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAA length\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eMolecular Weight\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eTheoretical PI\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eInstability index\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eAliphatic index\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003eGravy\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eLocalization\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eOchotona princeps\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e233\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e25476.85\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e6.60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e58.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e77.47\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e-0.289\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eMitochondria\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCavia porcellus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e200\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e22284.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e6.12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e61.85\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e68.25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e-0.562\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNuclear\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLynx Canadensis\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e205\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e22720.55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e6.23\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e65.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e67.61\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e-0.524\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eMitochondria\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eBos Taurus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e204\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e22679.34\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5.77\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e59.53\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e69.36\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e-0.597\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eMitochondria\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eOvis aries\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e201\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e22334.03\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e6.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e63.46\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e70.40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e-0.568\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNuclear\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCamelus ferus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e201\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e22410.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e6.09\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e65.52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e70.85\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e-0.551\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNuclear\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCapra hircus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e201\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e22349.00\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e6.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e62.71\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e68.46\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e-0.604\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNuclear\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003ePteropus alecto\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e207\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e22853.71\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e6.32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e73.49\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e66.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e-0.549\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNuclear\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eHomo sapiens\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e205\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e22782.52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5.98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e62.82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e68.54\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e-0.567\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNuclear\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eGorilla gorilla\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e205\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e22782.52\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5.98\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e62.82\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e68.54\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e-0.567\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNuclear\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eErinaceus europaeus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e199\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e22032.94\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e6.08\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e68.37\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e75.43\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e-0.438\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNuclear \u0026amp;\u003c/p\u003e \u003cp\u003eMitochondria\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eSuncus etruscus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e209\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e22988.85\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e6.22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e69.20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e65.84\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e-0.492\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003eNuclear \u0026amp;\u003c/p\u003e \u003cp\u003eMitochondria\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec13\" class=\"Section2\"\u003e \u003ch2\u003ePhylogenetic analysis\u003c/h2\u003e \u003cp\u003eWithin the evolutionary tree that was constructed with MEGA 11, five distinct groups, or clades, emerged. Figure\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003e illustrates quite clearly that members of clade 1, represented by the blue color, \u003cem\u003eS. etruscus\u003c/em\u003e and \u003cem\u003eE. europaeus\u003c/em\u003e, have descended from the same ancestor and are closely related to one another. In the second group, represented by the color red, \u003cem\u003eH. sapiens\u003c/em\u003e and \u003cem\u003eG. gorillas\u003c/em\u003e are closely related to one another since they share a common ancestor. \u003cem\u003eC. porcellus\u003c/em\u003e, \u003cem\u003eL. canadensis\u003c/em\u003e, and \u003cem\u003eO. princeps\u003c/em\u003e are the three species that originated from the same ancestor and make up the third clade, which is colored peach. All four of these species, \u003cem\u003eB. taurus\u003c/em\u003e, \u003cem\u003eO. aries, C. ferus, and C. hircus\u003c/em\u003e, belong to the same clade and are closely related to one another. \u003cem\u003eP. alecto\u003c/em\u003e is not closely related to any other organism and is depicted as a single entity in the phylogenetic tree even though it is located in clade 5, which is denoted by the branch color purple.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec14\" class=\"Section2\"\u003e \u003ch2\u003eMotif analysis\u003c/h2\u003e \u003cp\u003eThe investigation that was carried out with the assistance of the MEME tool revealed that motifs 1 through 7 were existing in the protein sequences of all species with the exception of \u003cem\u003eE. europaeus\u003c/em\u003e. Only the protein sequences of two different species, namely \u003cem\u003eS. etruscus\u003c/em\u003e and \u003cem\u003eO. princeps\u003c/em\u003e, contained the Motif 8 pattern. Two more species, namely \u003cem\u003eP. alecto\u003c/em\u003e and \u003cem\u003eO. princeps\u003c/em\u003e, were identified to have a motif 9 in their DNA. Motif 10 was unique to \u003cem\u003eO. princeps\u003c/em\u003e, as illustrated in Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e, and was only identified in that species.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec15\" class=\"Section2\"\u003e \u003ch2\u003eDomain analysis\u003c/h2\u003e \u003cp\u003eAccording to the results of the InterProScan study, all of the proteins from the species that were selected are members of the same homologous family called \"Homologous super family HSP20_like_Chaperson,\" having the accession number IPR008978. The accession numbers (IPR002068 and IPR037876) for both of the DNA building domains known as \"ACD_HSPB1 and A Crystalline\" for each species as shown in Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003ePositions of homologous super families and domains present in the selected organisms\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"4\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOrganism Name\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eHomologous Super family\u003c/p\u003e \u003cp\u003eHSP20_like_Chaperone\u003c/p\u003e \u003cp\u003eIPR008978\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eDomain 1\u003c/p\u003e \u003cp\u003eACD_HSPB1\u003c/p\u003e \u003cp\u003eIPR037876\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eDomain 2\u003c/p\u003e \u003cp\u003eA Crystalline\u003c/p\u003e \u003cp\u003eIPR002068\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eOchotona princeps\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e69\u0026ndash;189\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e83\u0026ndash;168\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e75\u0026ndash;183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCavia porcellus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e62\u0026ndash;187\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e79\u0026ndash;164\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e71\u0026ndash;179\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLynx canadensis\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e69\u0026ndash;191\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e76\u0026ndash;184\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e84\u0026ndash;169\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eBos Taurus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e67\u0026ndash;187\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e72\u0026ndash;180\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e80\u0026ndash;165\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eOvis aries\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e67\u0026ndash;187\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e72\u0026ndash;180\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e80\u0026ndash;165\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCamelus ferus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e65\u0026ndash;187\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e80\u0026ndash;165\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e72\u0026ndash;180\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCapra hircus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e67\u0026ndash;187\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e72\u0026ndash;180\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e80\u0026ndash;165\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003ePteropus alecto\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e72\u0026ndash;193\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e78\u0026ndash;186\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e86\u0026ndash;171\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eHomo sapiens\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e69\u0026ndash;197\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e84\u0026ndash;169\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e76\u0026ndash;184\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eGorilla gorilla\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e69\u0026ndash;197\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e84\u0026ndash;169\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e76\u0026ndash;184\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eErinaceus europaeus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e69\u0026ndash;190\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e83\u0026ndash;168\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e75\u0026ndash;183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eSuncus etruscus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e69\u0026ndash;190\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e83\u0026ndash;168\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e75\u0026ndash;183\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec16\" class=\"Section2\"\u003e \u003ch2\u003eGene structure analysis\u003c/h2\u003e \u003cp\u003eThe analysis of upstream, downstream, intron, and CDS regions was conducted using GSDS for each species. The term upstream and downstream refer to the relative positions of the genetic information within RNA or DNA, respectively. Specifically, downstream refers to the 3' end of the coding strand, while upstream refers to the 5' end of the coding strand. Both \u003cem\u003eB. taurus\u003c/em\u003e and \u003cem\u003eO. princeps\u003c/em\u003e possess all four components, upstream, downstream intron and CDS regions. Other species, including \u003cem\u003eC. porcellus, L. canadensis, O. aries, C. ferus, C. hircus, P. alecto, H. sapiens, G. gorilla, E. europaeus, and S. etruscus\u003c/em\u003e only possess intron and CDS regions of the gene.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec17\" class=\"Section2\"\u003e \u003ch2\u003ePercentage identity and similarity of HSPB1 Gene\u003c/h2\u003e \u003cp\u003eThe percentage of amino acids that are identical to one another and the identity matrix of amino acids are crucial factors in predicting evolutionary patterns and distinguishing different species. Tables\u0026nbsp;\u003cspan refid=\"Tab4\" class=\"InternalRef\"\u003e4\u003c/span\u003e and \u003cspan refid=\"Tab5\" class=\"InternalRef\"\u003e5\u003c/span\u003e present the the identity percentage and the similarity percentage of various species, respectively. A higher identity and similarity percentage between species typically indicates a closer relationship, suggesting that their structure and functions are are more likely to be comparable. In particular, sequences with a similarity percentage of approximately 70% are indicative of shared homology, functional similarity, and highly conserved regions HSPB1 gene. This gene plays a crucial role in cellular stress response and protein maintenance. The pair of species exhibiting a similarity of around 70% in their HSPB1 gene likely share similar functions and evolutionary histories.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab4\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 4\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe percentage of amino acids identical across different organisms\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"13\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c12\" colnum=\"12\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c13\" colnum=\"13\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eOchotona princeps\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eCavia porcellus\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003eLynx Canadensis\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003eBos taurus\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cem\u003eOvis aries\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cem\u003eCamelus ferus\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cem\u003eCapra hircus\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cem\u003ePteropus alecto\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cem\u003eHomo sapiens\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c11\"\u003e \u003cp\u003e\u003cem\u003eGorilla gorilla\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c12\"\u003e \u003cp\u003e\u003cem\u003eErinaceus europaeus\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c13\"\u003e \u003cp\u003e\u003cem\u003eSuncus etruscus\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eOchotona princeps\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e76.56%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e81.58%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e75.31%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e76.98%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e76.15%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e76.56%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e75.73%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e76.15%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e76.15%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e71.54%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e75.31%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCavia porcellus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e76.56%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e92.46%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e88.28%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e90.79%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e92.05%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e91.21%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e87.44%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e88.28%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e88.28%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e81.58%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e86.19%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLynx canadensis\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e81.58%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e92.46%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e89.12%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e92.05%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e91.21%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e91.63%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e90.37%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e90.37%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e90.37%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e83.68%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e89.53%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eBos taurus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e75.31%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e88.28%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e89.12%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e96.65%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e93.3%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e96.23%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e85.35%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e86.19%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e86.19%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e79.91%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e82.84%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eOvis aries\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e76.98%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e90.79%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e92.05%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e96.65%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e95.81%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e99.58%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e87.86%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e89.12%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e89.12%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e82%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e85.77%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCamelus ferus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e76.15%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e92.05%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e91.21%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e93.3%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e95.81%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e95.39%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e89.53%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e87.86%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e87.86%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e82.42%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e85.35%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCapra hircus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e76.56%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e91.21%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e91.63%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e96.23%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e99.58%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e95.39%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e87.44%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e89.53%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e89.53%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e81.58%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e86.19%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003ePteropus alecto\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e75.73%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e87.44%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e90.37%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e85.35%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e87.86%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e89.53%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e87.44%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e89.12%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e89.12%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e83.68%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e89.12%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eHomo sapiens\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e76.15%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e88.28%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e90.37%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e86.19%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e89.12%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e87.86%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e89.53%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e89.12%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e82.42%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e88.7%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eGorilla gorilla\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e76.15%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e88.28%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e90.37%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e86.19%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e89.12%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e87.86%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e89.53%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e89.12%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e82.42%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e88.7%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eErinaceus europaeus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e71.54%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e81.58%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e83.68%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e79.91%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e82%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e82.42%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e81.58%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e83.68%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e82.42%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e82.42%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e82.84%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eSuncus etruscus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e75.31%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e86.19%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e89.53%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e82.84%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e85.77%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e85.35%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e86.19%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e89.12%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e88.7%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e88.7%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e82.84%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab5\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 5\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe Percentage of amino acids similar across different organisms\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"13\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c11\" colnum=\"11\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c12\" colnum=\"12\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c13\" colnum=\"13\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cem\u003eOchotona princeps\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e\u003cem\u003eCavia porcellus\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e\u003cem\u003eLynx canadensis\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e\u003cem\u003eBos taurus\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e\u003cem\u003eOvis aries\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cem\u003eCamelus ferus\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cem\u003eCapra hircus\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cem\u003ePteropus alecto\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\"\u003e \u003cp\u003e\u003cem\u003eHomo sapiens\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c11\"\u003e \u003cp\u003e\u003cem\u003eGorilla gorilla\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c12\"\u003e \u003cp\u003e\u003cem\u003eErinaceus europaeus\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c13\"\u003e \u003cp\u003e\u003cem\u003eSuncus etruscus\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eOchotona princeps\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e75.73%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e80.33%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e74.47%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e75.73%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e75.73%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e75.31%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e76.56%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e75.73%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e75.73%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e71.12%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e76.56%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCavia porcellus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e75.73%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e78.66%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e74.89%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e76.15%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e78.24%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e76.56%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e75.73%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e75.73%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e75.73%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e69.45%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e75.31%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eLynx canadensis\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e80.33%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e78.66%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e76.15%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e77.82%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e77.82%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e77.4%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e78.24%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e77.4%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e77.4%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e71.12%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e77.82%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eBos taurus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e74.47%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e74.89%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e76.15%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e82%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e79.49%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e81.58%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e74.89%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e74.47%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e74.47%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e68.2%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e73.22%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eOvis aries\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e75.73%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e76.15%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e77.82%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e82%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e80.75%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e83.68%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e76.15%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e76.15%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e76.15%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e69.03%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e74.89%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCamelus ferus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e75.73%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e78.24%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e77.82%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e79.49%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e80.75%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e80.33%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e76.98%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e75.73%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e75.73%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e69.45%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e75.31%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eCapra hircus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e75.31%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e76.56%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e77.4%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e81.58%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e83.68%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e80.33%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e75.73%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e76.56%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e76.56%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e68.61%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e75.31%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003ePteropus alecto\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e76.56%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e75.73%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e78.24%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e74.89%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e76.15%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e76.98%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e75.73%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e78.24%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e78.24%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e72.38%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e78.66%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eHomo sapiens\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e75.73%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e75.73%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e77.4%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e74.47%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e76.15%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e75.73%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e76.56%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e78.24%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e85.77%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e70.71%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e78.66%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eGorilla gorilla\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e75.73%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e75.73%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e77.4%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e74.47%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e76.15%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e75.73%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e76.56%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e78.24%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e85.77%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e70.71%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e78.66%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eErinaceus europaeus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e71.12%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e69.45%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e71.12%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e68.2%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e69.03%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e69.45%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e68.61%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e72.38%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e70.71%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e70.71%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e73.22%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eSuncus etruscus\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e76.56%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e75.31%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e77.82%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e73.22%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e74.89%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e75.31%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e \u003cp\u003e75.31%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e78.66%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e \u003cp\u003e78.66%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c11\"\u003e \u003cp\u003e78.66%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c12\"\u003e \u003cp\u003e73.22%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c13\"\u003e \u003cp\u003e100%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec18\" class=\"Section2\"\u003e \u003ch2\u003eStructural analysis\u003c/h2\u003e \u003cp\u003eGORIV tool was used to make the prediction about the secondary structure of the HSPB1 gene for the 12 organisms that were chosen. According to Table \u003cspan refid=\"Tab6\" class=\"InternalRef\"\u003e6\u003c/span\u003e, the HSPB1 gene has an alpha helix, a random coil, and extended strands as part of its secondary structure. The HSPB1 gene has an alpha helix, a random coil, and extended strands as part of its secondary structure. The \u003cem\u003eL. canadensis\u003c/em\u003e alpha helix takes up a total of 26.83 percent of the structure. The portion of the structure that is occupied by the alpha helix, the least by \u003cem\u003eH. sapiens\u003c/em\u003e and \u003cem\u003eG. gorilla\u003c/em\u003e is 13.66 percent. In addition, the extended strand for the \u003cem\u003eB. taurus\u003c/em\u003e was the longest at 17.75%, while the extended strand for the \u003cem\u003eC. porcellus\u003c/em\u003e was the shortest at 11.00%. On the other hand, random coils make up 73.17% of the structure of \u003cem\u003eH. sapiens\u003c/em\u003e and \u003cem\u003eG. gorilla\u003c/em\u003e, which is the largest percentage of random coils among other creatures. For HSPB1 protein sequence, a 12, 3D structure templates were created, and all of the models were successfully constructed from Phyre2 (Fig.\u0026nbsp;\u003cspan refid=\"Fig6\" class=\"InternalRef\"\u003e6\u003c/span\u003e). The 3D structure of HSPB1 protein revealed details about their folding, stability, and domain interactions.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab6\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 6\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eAll the criteria and percentages for the secondary structure of the organisms\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOrganism Name\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eAlpha helix\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003eBeta Bridge\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eBeta Turn\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eExtended strand\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eRandom Coil\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eOchotona princeps\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e26.61%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e14.16%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e59.23%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eCavia porcellus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e21.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e11.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e68.00%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eLynx Canadensis\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e26.83%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e12.20%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e60.98%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eBos Taurus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e21.08%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e17.75%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e61.27%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eOvis aries\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e20.90%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e14.93%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e64.18%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eCamelus ferus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e19.90%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e16.40%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e63.68%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eCapra hircus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e20.90%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e12.94%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e66.17%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003ePteropus Alecto\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e18.36%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e13.53%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e68.12%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eHomo sapiens\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e13.66%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e13.17%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e73.17%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eGorilla gorilla\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e13.66%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e13.17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e73.17%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eErinaceus europaeus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e16.58%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e17.59%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e65.83%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cem\u003eSuncus etruscus\u003c/em\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e18.66%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e0.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0.00%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e12.44%\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e68.90%\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003c/div\u003e"},{"header":"Discussion","content":"\u003cp\u003eHeat shock protein 27, also known as HSPB1, is a member of the family of proteins known as small heat shock proteins, or sHSPs. This protein contributes to the development of a stable state during times of high stress, such as when the body is subjected to heat shock or oxidative damage [20]. The findings of this study were derived from a computational analysis of the HSPB1 protein found in a variety of species. During this computational analysis, a number of analyses were carried out in order to identify Domains, multiple sequence alignment, physiochemical properties, motifs, quaternary structure, secondary structure parameters, protein interaction with its functional proteins in a variety of organisms, similarity and identity matrix, and the Position and composition of conserved elements, introns, exons in protein. \u003c/p\u003e\n\u003cp\u003eA gene family\u0026apos;s genetic characterization can reveal details about the genes\u0026apos; origins, duplication events, and diversification throughout their evolutionary history. The conserved regions, which are crucial for protein structure and functional studies, can be found using multiple sequence alignment (MSA) [21]. In nucleus majority of the proteins were found, while the mitochondria only contained a small percentage. Except for \u003cem\u003eB. taurus\u003c/em\u003e and \u003cem\u003eO. princeps\u003c/em\u003e, all creatures had just CDS parts, downstream and upstream in their protein sequences. These two species have CDS parts, Introns, downstream and upstream in their protein sequences. When referring to the relative placement of the genetic code in DNA or RNA, the terms \u0026quot;downstream\u0026quot; and \u0026quot;upstream\u0026quot; are both used. Upstream is towards the 5\u0026apos; end of the coding strand of the protein that is now being analyzed, while downstream is towards the 3\u0026apos; end of the coding strand. Both ACD_HSPB1 IPR037876 and A Crystalline IPR002068 were identified as the domains, present in each and every organism. The homologous superfamily found in all organisms was designated as IPR008978 Hsp20. The motif from 1 to 7 was found in the protein sequences of all organisms, with the exception of \u003cem\u003eE. europaeus\u003c/em\u003e. Only the protein sequences of two different organisms, namely \u003cem\u003eS. etruscus \u003c/em\u003eand\u003cem\u003e O. princeps\u003c/em\u003e, contained the Motif 8 pattern. Two more organisms, namely \u003cem\u003eP. alecto\u003c/em\u003e and \u003cem\u003eO. princeps\u003c/em\u003e, were found to have a motif 9 in their DNA. While Motif 10 was unique to \u003cem\u003eO. princeps\u003c/em\u003e, it was detected in both species. Furthermore, the varied patterns of the introns and downstream/upstream untranslated regions (UTRs) in the gene structure suggest that it may be due to the inclusion and removal of retroposons [22]. A prior study [23] was also responsible for collecting amino acids, determining % similarity and identity, phylogenetic relationship, determining physiochemical qualities, and identifying motifs.\u003c/p\u003e\n\u003cp\u003eThe phylogenetic tree revealed the presence of five distinct clades. In clade 1(represented by blue color), the species \u003cem\u003eS. etruscus\u003c/em\u003e and \u003cem\u003eE. europaeus\u003c/em\u003e share common ancestor, indicating their close evolutionary relationship. In the second clade (marked in red), \u003cem\u003eH. sapiens \u003c/em\u003eand\u003cem\u003e G. gorillas\u003c/em\u003e are closely related, as evidenced by to their shared common ancestor. The third clade, colored peach, consists of \u003cem\u003eC. porcellus, L. canadensis, \u003c/em\u003eand\u003cem\u003e O. princeps, \u003c/em\u003ewhich all originated from a common ancestor, highlighting their evolutionary proximity. The fourth clade includes \u003cem\u003eB. taurus\u003c/em\u003e, \u003cem\u003eO. aries, C. ferus\u003c/em\u003e, and\u003cem\u003e C. hircus\u003c/em\u003e, which are all closely related, stemming from a shared lineage. Lastly,\u003cem\u003e P. alecto\u003c/em\u003e is depicted as a distinct entity in clade 5 (marked in purple), isolated from other species, despite being located within this clade. The phylogenetic analysis also revealed a high degree of sequence homology among the species, reinforcing their evolutionary relationships as observed in other studies [24]. \u003c/p\u003e\n\u003cp\u003eFurthermore, assessing the physiochemical characteristics of the gene encoding the gene is necessary to comprehend the protein structure and function of the gene family [25]. According to the findings, every organism has a greater tendency towards hydrophilicity. The aliphatic index is a measure that may be used to evaluate the thermal stability of proteins; in this case, its value was greater than 50% for every protein. This suggests that aliphatic side amino acids covered over half of the volume of these proteins. In addition, GRAVY values that are lower than zero indicate that the hydrophilicity of these proteins is greater than that of their surroundings. The fact that all of the organisms have theoretical PI values that are lower than 7 demonstrates that they are all acidic in their natural state. Because the instability index for every creature was greater than 40, it may be deduced that every protein in every organism, with one exception, is unstable. Similarly, since GRAVY indicates a protein\u0026apos;s hydrophilicity (GRAVY \u0026lt;0) or hydrophobicity (GRAVY \u0026gt;0), assessing it would aid in a better understanding of its tertiary structure or shape [26]. Similar genetic characterization was done in previous studies including the assembly of amino acids and nucleotides, the computation of similarity and identity matrices, the assessment of physiochemical properties, the analysis of evolutionary relationships, the characterization of structural features, and the generation of a hydrophobicity plot [27]. Hence in recent years, determining and quantifying the evolutionary processes that contributed to genetic variety has emerged as an exciting area of research.\u003c/p\u003e\n\u003cp\u003eThe secondry structure of the HSPB1 gene in 12 species was predicted using GORIV tool, indicating the presence of alpha helices, random coils and extended strands. \u003cem\u003eL. canadensis\u003c/em\u003e showed the highest alpha helix content, while \u003cem\u003eH. sapiens\u003c/em\u003e and\u003cem\u003e G. gorillas \u003c/em\u003ehad the largest proportion of random coils. The three-dimensional models generated for the HSPB1 protein provided insights into its folding. Stability and domain interactions. These findings align with the previous studies on protein structure-function relationship and emphasize the evolutionary conservation of the HSPB1 gene across the species [28],[29].\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eThis study provides comprehensive insights into the evolutionary history, functional motifs, physiochemical characteristics and structural traits of the HSPB1 gene across various species. The results revealed the vital role of HSPB1 proteins in cellular processes, emphasizinf their acidic nature and high sequence conservation. Phgylogenetic analysis highlighted the evolutionary relationship among species, depicting their shared ancestry, The GRAVY values pointed to the hydrophilic nature of these proteins while the aliphatic index indicated their heat stability, further emphasizing their functional significance. The study also unveiled conserved motifs and homologous superfamily memberships which underscore the functional conservation of HSPB1 gene across the species. Variations in genetic organization, like the presence of distinct upstream and downstream regions in certain species reflect the complexity of gene regulation. Furthermore, the presence of conserved regions within the supports the notion of functional similarity across the species. The identity and similarity matrices confirmed that the species with higher similarity percentages in the HSPB1 gene likely shre common functions and evolutionary backgrounds. The analysis of secondary structure, including the distribution of alpha helices, random coils and extended strands, provided valuable insights on the protein folding, stability and domain interactions, the three-dimensional structure \u0026nbsp;of HSPB1 protein offered further insights into its functional architecture. This work identifies HSPB1 as functionally significant and underscores the need for additional research to fully comprehend the function and mechanism of HSPs.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eConflict of Interest:\u0026nbsp;\u003c/strong\u003eAuthors have declared no conflict of interest.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAcknowledgements:\u003c/strong\u003e None\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eData Availability Statement:\u0026nbsp;\u003c/strong\u003eAll the data is included in the manuscript.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors’ Contribution:\u0026nbsp;\u003c/strong\u003eM.S wrote original draft, S.P conceived main idea, A.K and M.A.M performed analysis, S.J and M. H wrote methodology, S.S and M.T prepared tables, M.F.K revised the manuscript and project administration.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eMymrikov, E.V., A.S. Seit-Nebi, and N.B. 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Shafique, \u0026ldquo;Molecular characterization of doublesex and Mab-3 (DMRT) gene family in Ctenopharyngodon idella (grass carp),\u0026rdquo; \u003cem\u003eJ Appl Genet\u003c/em\u003e, pp. 1\u0026ndash;12, 2024.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"Department of Zoology, The Government Sadiq College Women University, 63100, Bahawalpur, Punjab, Pakistan.","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"Domain, Gene structure, Genomics, HSPB1, Motif, Phylogeny","lastPublishedDoi":"10.21203/rs.3.rs-6417248/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6417248/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eHeat Shock Protein Beta-1 (HSPB1), a molecular chaperone crucial for celluar response and proteostasis, exhibits evolutionary conservation woth potential lineage-specific adaptations in placental mammals, warranting detailed comparative genomic investigation. The study investigated the characteristics, evolutionary links, motifs, secondary structure, and genetic organization of Heat Shock Protein Beta-1 \u003cb\u003e(\u003c/b\u003eHSPB1) protein across twelve distinct mammals. Significant sequence conservation was identified using multiple sequence alignments (MSA), with over 70% identity in specific areas among the chosen organisms. Physiochemical analysis revealed that all species\u0026rsquo; protein sequences exhibited an acidic nature, while instability indices indicated inherent protein instability. The GRAVY analysis referred to hydrophilic properties, while the aliphatic index showed heat stability. Phylogenetic analysis revealed five distinct clades, corresponding to major placental mammals\u0026rsquo; groups (e.g. \u003cem\u003eHomo sapiens, Bos Taurus\u003c/em\u003e), which underscores deep evolutionary divergences and conserved stress-response adaptations across lineages. Motif analysis revealed distinctive patterns in several species, and InterProScan results revealed membership in the \"Homologous superfamily HSP20_like_Chapserson\" family. An examination of the genetic organization indicated differences among organisms in the upstream, downstream, intron, and CDS regions, and the presence of conserved regions suggested their identity and similarity matrices. The current study conducted a computational approach and supporting evidence that HSPB1 is a novel heat shock responsive protein identified in placental mammals The current study findings provide a foundational framework delving into HSPB1 evolutionary and lineage-specific diversification, offering valuable insights into stress adaptation mechanisms and their implications for biomedical or evolutionary studies in mammals.\u003c/p\u003e","manuscriptTitle":"Identification, Genomic Characterization, and Phylogenetic Relationship of the Heat Shock Protein Beta-1 (HSPB1) in Placental Mammals","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-04-11 03:58:10","doi":"10.21203/rs.3.rs-6417248/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"c8ad15c1-2893-4a40-a230-0b3103878503","owner":[],"postedDate":"April 11th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[{"id":46952346,"name":"Animal Science"}],"tags":[],"updatedAt":"2025-04-11T03:58:10+00:00","versionOfRecord":[],"versionCreatedAt":"2025-04-11 03:58:10","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6417248","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6417248","identity":"rs-6417248","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}