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In our attempts to extend the utility of photoreactive reporters to map the molecular characteristics of vital membrane proteins, we carried out site\u2010selective incorporation of 7\u2010fluoro\u2010indole in the human mitochondrial outer membrane protein VDAC2 (voltage\u2010dependent anion channel isoform 2), with the aim of generating Trp\u2212Phe\/Tyr cross\u2010links. Prolonged irradiation at 282\u2005nm provided us with a surprisingly unusual fluorophore that displayed sizably red\u2010shifted excitation (\u03bb<jats:sub>ex\u2010max<\/jats:sub>=280\u2005nm\u2192360\u2005nm) and emission (\u03bb<jats:sub>em\u2010max<\/jats:sub>=330\u2005nm\u2192430\u2005nm) spectra that was reversible with organic solvents. By measuring the kinetics of the photo\u2010activated cross\u2010linking with a library of hVDAC2 variants, we demonstrate that formation of this unusual fluorophore is kinetically retarded, independent of tryptophan, and is site\u2010specific. Using other membrane (Tom40 and Sam50) and cytosolic (MscR and DNA Pol I) proteins, we additionally show that formation of this fluorophore is protein\u2010independent. Our findings reveal the photoradical\u2010mediated accumulation of reversible tyrosine cross\u2010links, with unusual fluorescent properties. Our findings have immediate applications in protein biochemistry and UV\u2010mediated protein aggregation and cellular damage, opening avenues for formulating therapeutics that prolong cell viability in humans.<\/jats:p>","DOI":"10.1002\/cbic.202300380","type":"journal-article","created":{"date-parts":[[2023,5,26]],"date-time":"2023-05-26T05:22:58Z","timestamp":1685078578000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["Photoradical\u2010Mediated Catalyst\u2010Independent Protein Cross\u2010Link with Unusual Fluorescence Properties"],"prefix":"10.1002","volume":"24","author":[{"given":"Jinam Ravindra","family":"Bora","sequence":"first","affiliation":[{"name":"Molecular Biophysics Laboratory Department of Biological Sciences Indian Institute of Science Education and Research  Bhopal Bypass Road, Bhauri Bhopal 462066 Madhya Pradesh 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