Direct Screening for Phosphatase Activity by Turnover-Based Capture of Protein Catalysts

• Published in: Angewandte Chemie International Edition Vol. 41; no. 5; pp. 775 - 777
• Main Authors: Betley, Jason R., Cesaro-Tadic, Sandro, Mekhalfia, Abdelaziz, Rickard, James H., Denham, Hazel, Partridge, Lynda J., Plückthun, Andreas, Blackburn, G. Michael
• Format: Journal Article
• Language: English
• Published: Weinheim WILEY-VCH Verlag GmbH 01.03.2002; WILEY‐VCH Verlag GmbH; Wiley
• Subjects: Biosensing Techniques - methods; biosensors; Catalysis; catalytic antibodies; Chemistry; Chemistry, Multidisciplinary; enzyme catalysis; Enzyme Inhibitors - chemical synthesis; Enzyme Inhibitors - chemistry; Humans; inhibitors; Kinetics; phosphatases; Phosphoric Monoester Hydrolases - analysis; Phosphoric Monoester Hydrolases - antagonists & inhibitors; Phosphoric Monoester Hydrolases - isolation & purification; Physical Sciences; Science & Technology; Serum Albumin, Bovine - chemistry; INACTIVATION; inhibitors; MECHANISM; catalytic antibodies; enzyme catalysis; biosensors; PROSTATIC ACID-PHOSPHATASE; 2-IMINOTHIOLANE; SELECTION; phosphatases
• ISSN: 1433-7851; 1521-3773
• DOI: 10.1002/1521-3773(20020301)41:5<775::AID-ANIE775>3.0.CO;2-F

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.