Insights into rhodopsin molecular evolution from mice with “humanized” Phe-88 to Leu substitution

Insights into rhodopsin molecular evolution from mice with “humanized” Phe-88 to Leu substitution | 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 Article Insights into rhodopsin molecular evolution from mice with “humanized” Phe-88 to Leu substitution Feifei Wang, Alexander V. Kolesnikov, Shinya Sato, Aneal Singh, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7606538/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 26 Jan, 2026 Read the published version in Scientific Reports → Version 1 posted 12 You are reading this latest preprint version Abstract The function of rod photoreceptors as dim light photon detectors depends critically on the molecular properties of their visual pigment, rhodopsin. The structure of rhodopsin has evolved under selective pressure to light conditions of different spectral composition and overall intensity. One notable example is the switch of mammalian species from nocturnal to diurnal environments. Comparison of the rhodopsins of the nocturnal mouse and the diurnal human reveals high sequence similarity, with only 18 distinct amino acids. Here, we examined the role of one of these, mouse phenylalanine (F) vs. human leucine (L) at position 88, in modulating the molecular properties of rhodopsin and the function of rods by generating F88L rhodopsin knock-in mouse. Our detailed in vitro analysis of the physicochemical properties of this mutant F88L rhodopsin showed a higher conformational stability and more efficient chromophore regeneration compared to the WT mouse pigment. We also found that the decay of metarhodopsin II in the F88L mutant occurred significantly faster than in the WT. However, despite these molecular changes, the visual function of knock-in mutant mice carrying the F88L mutation was not significantly altered by this amino acid change. These findings demonstrate the role of the F88L evolutionary switch in enhancing the stability and regeneration of rhodopsin towards visual function in diurnal human rods over nocturnal mouse rods. Our results provide new insights into the molecular evolution of rhodopsin in vertebrates. Biological sciences/Biochemistry Biological sciences/Biophysics Biological sciences/Neuroscience Rhodopsin Phototransduction Molecular evolution Nocturnal vision Diurnal vision Full Text Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 26 Jan, 2026 Read the published version in Scientific Reports → Version 1 posted Editorial decision: Revision requested 17 Oct, 2025 Reviews received at journal 17 Oct, 2025 Reviewers agreed at journal 16 Oct, 2025 Reviews received at journal 06 Oct, 2025 Reviewers agreed at journal 03 Oct, 2025 Reviewers agreed at journal 02 Oct, 2025 Reviewers agreed at journal 01 Oct, 2025 Reviewers invited by journal 01 Oct, 2025 Editor assigned by journal 01 Oct, 2025 Editor invited by journal 01 Oct, 2025 Submission checks completed at journal 30 Sep, 2025 First submitted to journal 30 Sep, 2025 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. 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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-7606538","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":528339307,"identity":"192e0d6d-f9cc-4495-a5b4-b1940681f1e3","order_by":0,"name":"Feifei Wang","email":"","orcid":"","institution":"Universitat Politècnica de Catalunya-Barcelona Tech","correspondingAuthor":false,"prefix":"","firstName":"Feifei","middleName":"","lastName":"Wang","suffix":""},{"id":528339308,"identity":"4dc42044-f2a4-459c-9653-b10605352f50","order_by":1,"name":"Alexander V. 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