Proteolysis-assisted cyclization facilitates site-centric target deconvolution of isothiocyanates

Proteolysis-assisted cyclization facilitates site-centric target deconvolution of isothiocyanates | 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 Proteolysis-assisted cyclization facilitates site-centric target deconvolution of isothiocyanates Jing Yang, Jing-Yang He, Caiping Tian, Ji-Xiang He, Qiang Li, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-5055115/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 Isothiocyanates (ITCs) are a unique class of electrophilic natural products that exert biological effects by reacting with proteinous cysteines to generate thionoacyl adducts. However, the identification of ITCs’ target sites is still an unmet task due to the high lability of such adducts. Here, we report an unexpected chemistry through which the ITC-protein adduct forms a stable N-terminal dihydrothiazole peptide adduct during proteolysis. This proteolysis-assisted cyclization (PAC) reaction can be harnessed for developing affinity-based and activity-based chemoproteomic methods to site-specifically profile targets of ITCs. Applying these methods not only add further complexity to the known poly-pharmacological landscape sulforaphane, the most studied ITC, but also permits expanding the ligandable cysteinome with a 55-member library of structurally diversified ITC-based fragments. Given the promising chemo-preventive and therapeutic effects of ITCs, the PAC-based chemoproteomic platform may lay the groundwork for elucidating their mechanisms of action and ultimately diversify cysteine targetability for drug discovery. Biological sciences/Chemical biology/Target identification Biological sciences/Chemical biology/Proteomics Full Text Additional Declarations There is NO Competing Interest. Supplementary Files SupplementaryData3R1.xlsx Supplementary Data 3 SupplementaryData5R1.xlsx Supplementary Data 5 SupplementaryData7R1.xlsx Supplementary Data 7 SupplementaryData2R1.xlsx Supplementary Data 2 SupplementaryData6R1.xlsx Supplementary Data 6 SupplementaryData8R1.xlsx Supplementary Data 8 SupplementaryData4R1.xlsx Supplementary Data 4 SupplementaryData1R1.xlsx Supplementary Data 1 SupplementaryInformationR2.pdf Supplementary Information 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. 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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-5055115","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Article","associatedPublications":[],"authors":[{"id":525986175,"identity":"bd6770ac-57be-4d89-81bb-974b80ac2a2c","order_by":0,"name":"Jing 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