Detection of an anhydride intermediate in the carboxypeptidase A catalyzed hydrolysis of a peptide substrate by solid state NMR spectroscopy and its mechanistic implication.
We have detected an anhydride intermediate in the CPA catalyzed proteolytic reaction of Gly-Tyr. It appears that since the zinc-bound water molecule which is believed to attack the scissile amide carbonyl carbon in the hydrolysis reaction is excluded by the N-terminal amino group of Gly-Tyr, the carboxylate of Glu-270 becomes to attack the amide bond to generate the anhydride intermediate.
Lee
H C
HC
Department of Chemistry, Pohang University of Science and Technology, Korea.
Ko
Y H
YH
Baek
S B
SB
Kim
D H
DH
D000812:Anhydrides / Q000032:analysis*; D002268:Carboxypeptidases / Q000378:metabolism*; D043422:Carboxypeptidases A; D002384:Catalysis; D004151:Dipeptides / Q000378:metabolism*; D006868:Hydrolysis; D009682:Magnetic Resonance Spectroscopy
D000812:Anhydrides; D004151:Dipeptides; C022013:glycyltyrosine; D002268:Carboxypeptidases; D043422:Carboxypeptidases A