title: Coumarin-based fluorogenic probes for no-wash protein labeling.
abstract: A fluorescent protein-labeling strategy was developed in which a protein of interest (POI) is genetically tagged with a short peptide sequence presenting two Cys residues that can selectively react with synthetic fluorogenic reagents. These fluorogens comprise a fluorophore and two maleimide groups that quench fluorescence until they both undergo thiol addition during the labeling reaction. Novel fluorogens were prepared and kinetically characterized to demonstrate the importance of a methoxy substituent on the maleimide in suppressing reactivity with glutathione, an intracellular thiol, while maintaining reactivity with the dithiol tag. This system allows the rapid and specific labeling of intracellular POIs.
authors.affiliation: Department of Chemistry, University of Ottawa, 10 Marie-Curie, Ottawa, ON K1N 6N5 (Canada).
authors.forename: Yingche
authors.initials: Y
authors.lastname: Chen
authors.forename: Christopher M
authors.initials: CM
authors.lastname: Clouthier
authors.forename: Kelvin
authors.initials: K
authors.lastname: Tsao
authors.forename: Miroslava
authors.initials: M
authors.lastname: Strmiskova
authors.forename: Hugo
authors.initials: H
authors.lastname: Lachance
authors.forename: Jeffrey W
authors.initials: JW
authors.lastname: Keillor
mesh_terms: D003374:Coumarins / Q000737:chemistry*; D005456:Fluorescent Dyes / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*
chemical_list: D003374:Coumarins; D005456:Fluorescent Dyes; D011506:Proteins; C030123:coumarin
keywords: bioorthogonal chemistry; fluorescent probes; glutathione; maleimide; protein labeling