Further studies on the activation of bovine pancreatic procarboxypeptidase A by trypsin.
Unlike the pancreatic endopeptidase zymogens, procarboxypeptidase A is activated very slowly in vitro. The activation proceeds through the removal of about 100 amino acids away from the N-terminus of the chain. The cleavage of the susceptible bond(s) in monomeric and aggregated forms of bovine procarboxypeptidase A by catalytic amounts of trypsin was found to be very fast. However, as in the case of the porcine zymogen, the expression of the carboxypeptidase activity was considerably delayed by the inhibitory effect of the activation peptide which remains bound to the enzyme molecule after the trypsin treatment of the zymogen. alpha-Carboxypeptidase A was mainly formed under the relatively mild conditions used, indicating that the Arg-1-Ala+1 bond is probably the first to be cleaved during in vitro activation. The bovine carboxypeptidase activity was immediately and reversibly expressed upon dimethylmaleylation of the activation mixtures. This expression does not require full dissociation of the enzyme-peptide complex but merely a suitable change in its quaternary structure resulting from a modification of some electrostatic interactions upon dimethylmaleylation. Separation of bovine carboxypeptidase A from its activation peptide was only achieved upon filtration of the dimethylmaleylated mixtures in the presence of 6 M urea. The bovine activation peptide contains at least 93 amino acids compared to the 94 amino acids found by other authors for the rat and porcine peptides and sequencing of the first 53 amino acids showed a 75-85% homology with the latter two peptides.
D000595:Amino Acid Sequence; D000818:Animals; D002268:Carboxypeptidases / Q000302:isolation & purification / Q000378:metabolism*; D043422:Carboxypeptidases A; D002417:Cattle; D004789:Enzyme Activation; D004792:Enzyme Precursors / Q000302:isolation & purification / Q000378:metabolism*; D007700:Kinetics; D008970:Molecular Weight; D010179:Pancreas / Q000201:enzymology*; D013552:Swine; D014357:Trypsin / Q000378:metabolism*
D004792:Enzyme Precursors; D002268:Carboxypeptidases; D043422:Carboxypeptidases A; D014357:Trypsin