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A Strategy for Functional Proteomic Analysis of Glycosidase Activity from Cell Lysates
Author(s) -
Vocadlo David J.,
Bertozzi Carolyn R.
Publication year - 2004
Publication title -
angewandte chemie international edition
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.831
H-Index - 550
eISSN - 1521-3773
pISSN - 1433-7851
DOI - 10.1002/anie.200454235
Subject(s) - glycoside hydrolase , chemistry , azide , covalent bond , steric effects , enzyme , nucleophile , adduct , biochemistry , stereochemistry , catalysis , organic chemistry
A multipurpose flag : The inactivator 1 covalently labels the catalytic nucleophiles of retaining β‐glycosidases to form species 2 . The small azide group allows labeling of enzymes with sterically congested active sites. Staudinger ligation of 2 with phosphine–FLAG yields adduct 3 , which can be used to detect and profile retaining β‐glycosidase activities in complex mixtures.