Premium
Direct Screening for Phosphatase Activity by Turnover‐Based Capture of Protein Catalysts
Author(s) -
Betley Jason R.,
CesaroTadic Sandro,
Mekhalfia Abdelaziz,
Rickard James H.,
Denham Hazel,
Partridge Lynda J.,
Plückthun Andreas,
Blackburn G. Michael
Publication year - 2002
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 - 10.1002/1521-3773(20020301)41:5<775::aid-anie775>3.0.co;2-f
Subject(s) - phosphate , hydrolysis , covalent bond , catalysis , chemistry , electrophile , phosphatase , disulfide bond , stereochemistry , biochemistry , enzyme , combinatorial chemistry , organic chemistry
Disulfide exchange leads to the attachment of phosphate ester 1 to a solid surface, which can then be used in turnover selection to screen a large library of proteins for phosphate monoester hydrolysis activity. This ester hydrolysis leads to formation of an electrophilic quinone methide 2 , which bonds covalently to the protein catalyst and links it to the surface for selection.