Thermodynamic coupling between folding correctors and the first of dimerized nucleotide binding domains in CFTR

Thermodynamic coupling between folding correctors and the first of dimerized nucleotide binding domains in CFTR | 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 Thermodynamic coupling between folding correctors and the first of dimerized nucleotide binding domains in CFTR Guangyu Wang This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6890276/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 29 Jul, 2025 Read the published version in ACS Bio & Med Chem Au → Version 1 posted You are reading this latest preprint version Abstract The most common cystic fibrosis mutation is the F508del mutation in the human cystic fibrosis transmembrane conductance regulator (hCFTR), which causes misfolding of the first of two nucleotide binding domains (NBD1/2), preventing Mg/ATP-dependent NBD dimerization for normal function. Although folding correctors elexacaftor/VX-445 and lumacaftor/VX-809 have been combined to correct the NBD1 misfolding, the exact correction pathway is still unknown. In this study, the constrained tertiary noncovalent interaction networks or the thermoring structures of dimerized NBD1 in hCFTR/E1371Q with or without F508del were analyzed to identify the weakest noncovalent bridge as the final posttranslational tertiary folding of dimerized NBD1 in response to folding correctors. These computational analyses suggested that hCFTR may primarily use cooperative folding between α- and β-subdomains in dimerized NBD1 as the last step upon the binding of the potentiator ivacaftor/VX-770. However, the binding of folding correctors may allosterically protect the α-subdomain from misfolding until subsequent core formation. This thermodynamic protective mechanism, unlike the chaperone-based one in cotranslational NBD1 folding, may restore posttranslational NBD1 folding for tight Mg/ATP-mediated NBD dimerization in the F508del mutation, and also potentially apply to treating other cystic fibrosis patients with rare mutations. General Biochemistry Biophysics Biochemical Research Methods Structural Biology Bioinformatics Computational Biology Systems Biology Chemical Biology Mathematical and Theoretical Biology Drug Discovery, Design, & Development Systems and Networking allosteric pathway cooperative folding digital biology energy landscape melting threshold noncovalent thermoring structure protective folding protein stability Full Text Additional Declarations The authors declare no competing interests. Supplementary Files SupportingInformationdimerizedNBD1andFoldingcorrectorsinhCFTRv1.0.pdf Cite Share Download PDF Status: Published Journal Publication published 29 Jul, 2025 Read the published version in ACS Bio & Med Chem Au → 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-6890276","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":471054581,"identity":"58b12fa9-875d-4ba8-aecb-5b9849a9acf6","order_by":0,"name":"Guangyu Wang","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA60lEQVRIiWNgGAWjYFADZsbGB0CKh494LezMhw1AWtiI18LPliYBoglqkXc/+0yCsc0uT96Zx6zya46dDBsD88NHN/BoMTyTbgbUklxseJjH7LbstmSgw9iMjXPwaWlIY5Ng3MacuLEZqEVyGzNQCw+bNF4t/c9AWurBWoolt9UT1iIvAbblcOJ8ZrY0xo/bDhPWYiDxjNki8d/xxA3MzIelGbcd52FjJuAX+f40xhsfzlQnzu8/2Pjx57Zqe3725oeP8dpygIFFIgHCYGDmAQkx41EOtqWBgfkDlMHA+IOA6lEwCkbBKBiZAACt70H8iuWZ5QAAAABJRU5ErkJggg==","orcid":"","institution":"","correspondingAuthor":true,"prefix":"","firstName":"Guangyu","middleName":"","lastName":"Wang","suffix":""}],"badges":[],"createdAt":"2025-06-13 18:27:47","currentVersionCode":1,"declarations":{"humanSubjects":false,"vertebrateSubjects":false,"conflictsOfInterestStatement":false,"humanSubjectEthicalGuidelines":false,"humanSubjectConsent":false,"humanSubjectClinicalTrial":false,"humanSubjectCaseReport":false,"vertebrateSubjectEthicalGuidelines":false},"doi":"10.21203/rs.3.rs-6890276/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6890276/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1021/acsbiomedchemau.5c00014","type":"published","date":"2025-07-30T00:00:00+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":88042947,"identity":"471be55e-540c-4dbf-8c4f-446f30709258","added_by":"auto","created_at":"2025-07-31 17:34:50","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":1040959,"visible":true,"origin":"","legend":"","description":"","filename":"ThermodynamiccouplingbetweenfoldingcorrectorsanddimerizedNBD1inhCFTRv1.0.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6890276/v1_covered_8f92b371-4168-4af6-bced-990dd87d7f87.pdf"},{"id":84759860,"identity":"af4fe3b5-38d7-4089-bfe6-33ea114878e6","added_by":"auto","created_at":"2025-06-17 05:41:27","extension":"pdf","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":420317,"visible":true,"origin":"","legend":"","description":"","filename":"SupportingInformationdimerizedNBD1andFoldingcorrectorsinhCFTRv1.0.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6890276/v1/ebfecfdccfcd869377345200.pdf"}],"financialInterests":"The authors declare no competing interests.","formattedTitle":"\u003cp\u003eThermodynamic coupling between folding correctors and the first of dimerized nucleotide binding domains in CFTR\u003c/p\u003e","fulltext":[],"fulltextSource":"","fullText":"","funders":[{"identity":"3dac0f0a-5f83-4eab-9561-20385d5ba241","identifier":"10.13039/100000968","name":"American Heart Association","awardNumber":"10SDG4120011","order_by":0}],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":false,"highlight":"","institution":"University of California, Davis","isAcceptedByJournal":true,"isAuthorSuppliedPdf":true,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":true,"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":"allosteric pathway, cooperative folding, digital biology, energy landscape, melting threshold, noncovalent thermoring structure, protective folding, protein stability","lastPublishedDoi":"10.21203/rs.3.rs-6890276/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6890276/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eThe most common cystic fibrosis mutation is the F508del mutation in the human cystic fibrosis transmembrane conductance regulator (hCFTR), which causes misfolding of the first of two nucleotide binding domains (NBD1/2), preventing Mg/ATP-dependent NBD dimerization for normal function. \u0026nbsp;Although folding correctors elexacaftor/VX-445 and lumacaftor/VX-809 have been combined to correct the NBD1 misfolding, the exact correction pathway is still unknown. In this study, the constrained tertiary noncovalent interaction networks or the thermoring structures of dimerized NBD1 in hCFTR/E1371Q with or without F508del were analyzed to identify the weakest noncovalent bridge as the final posttranslational tertiary folding of dimerized NBD1 in response to folding correctors. These computational analyses suggested that hCFTR may primarily use cooperative folding between α- and β-subdomains in dimerized NBD1 as the last step upon the binding of the potentiator ivacaftor/VX-770. \u0026nbsp;However, the binding of folding correctors may allosterically protect the α-subdomain from misfolding until subsequent core formation. This thermodynamic protective mechanism, unlike the chaperone-based one in cotranslational NBD1 folding, may restore posttranslational NBD1 folding for tight Mg/ATP-mediated NBD dimerization in the F508del mutation, and also potentially apply to treating other cystic fibrosis patients with rare mutations.\u003c/p\u003e","manuscriptTitle":"Thermodynamic coupling between folding correctors and the first of dimerized nucleotide binding domains in CFTR","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-06-17 05:41:22","doi":"10.21203/rs.3.rs-6890276/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":"43ee4efc-79ce-40f4-ace3-56b2fcffba09","owner":[],"postedDate":"June 17th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[{"id":50156681,"name":"General Biochemistry"},{"id":50156682,"name":"Biophysics"},{"id":50156683,"name":"Biochemical Research Methods"},{"id":50156684,"name":"Structural Biology"},{"id":50156685,"name":"Bioinformatics"},{"id":50156686,"name":"Computational Biology"},{"id":50156687,"name":"Systems Biology"},{"id":50156688,"name":"Chemical Biology"},{"id":50156689,"name":"Mathematical and Theoretical Biology"},{"id":50156690,"name":"Drug Discovery, Design, \u0026 Development"},{"id":50156691,"name":"Systems and Networking"}],"tags":[],"updatedAt":"2025-07-31T17:34:44+00:00","versionOfRecord":{"articleIdentity":"rs-6890276","link":"https://doi.org/10.1021/acsbiomedchemau.5c00014","journal":{"identity":"acs-bio-and-med-chem-au","isVorOnly":true,"title":"ACS Bio \u0026 Med Chem Au"},"publishedOn":"2025-07-30 00:00:00","publishedOnDateReadable":"July 30th, 2025"},"versionCreatedAt":"2025-06-17 05:41:22","video":"","vorDoi":"10.1021/acsbiomedchemau.5c00014","vorDoiUrl":"https://doi.org/10.1021/acsbiomedchemau.5c00014","workflowStages":[]},"version":"v1","identity":"rs-6890276","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6890276","identity":"rs-6890276","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}