title: A simple method for enhancing the bioorthogonality of cyclooctyne reagent.
abstract: The cross-reactivity between some cyclooctynes and thiols limits the bioorthogonality of the strain-promoted azide-alkyne cycloaddition reaction. We show that a low concentration of β-mercaptoethanol significantly reduces the undesirable side reaction between bicyclononyne (BCN) and cysteine and while preserving free cysteines. We site-specifically label a genetically-encoded azido group in the visual photoreceptor rhodopsin to demonstrate the utility of the strategy.
authors.affiliation: Laboratory of Chemical Biology and Signal Transduction, The Rockefeller University, 1230 York Ave., New York, NY 10065, USA. sakmar@rockefeller.edu hubert@rockefeller.edu.
authors.forename: He
authors.initials: H
authors.lastname: Tian
authors.affiliation: Laboratory of Chemical Biology and Signal Transduction, The Rockefeller University, 1230 York Ave., New York, NY 10065, USA. sakmar@rockefeller.edu hubert@rockefeller.edu and Department of Neurobiology, Care Sciences and Society, Division for Neurogeriatrics, Center for Alzheimer Research, Karolinska Institutet, 141 57 Huddinge, Sweden.
authors.forename: Thomas P
authors.initials: TP
authors.lastname: Sakmar
authors.affiliation: Laboratory of Chemical Biology and Signal Transduction, The Rockefeller University, 1230 York Ave., New York, NY 10065, USA. sakmar@rockefeller.edu hubert@rockefeller.edu.
authors.forename: Thomas
authors.initials: T
authors.lastname: Huber
mesh_terms: D000480:Alkynes / Q000737:chemistry*; D000818:Animals; D001386:Azides / Q000737:chemistry*; D002417:Cattle; D061565:Cycloaddition Reaction; D034242:Cyclooctanes / Q000737:chemistry*; D003545:Cysteine / Q000737:chemistry*; D008623:Mercaptoethanol / Q000737:chemistry*; D008958:Models, Molecular; D012243:Rhodopsin / Q000737:chemistry; D013438:Sulfhydryl Compounds / Q000737:chemistry
chemical_list: D000480:Alkynes; D001386:Azides; D034242:Cyclooctanes; D013438:Sulfhydryl Compounds; D008623:Mercaptoethanol; D012243:Rhodopsin; D003545:Cysteine