title: Two-step dissociation of bovine 6S procarboxypeptidase A by dimethylmaleylation.
abstract: Reversible condensation of the ternary complex form of bovine pancreatic procarboxypeptidase A with 2,3-dimethyl maleic anhydride was investigated at pH 9.0 and low concentration of reagent over the acylable amino groups. After subsequent modification of only a few lysyl residues, subunit III was found to have been released from the quaternary structure leading to the separation of an apparently native protein devoid of any contaminating subunit II, while dissociation of the remaining binary complex occurred upon further addition of the anhydride. This observation suggests that the electrostatic interactions existing between subunits I and III are more rapidly weakened than those between subunits I and II, probably because fewer lysyl residues are involved and/or there is greater accessibility to the chemical reagent . Although completely inactive on the specific substrates of trypsin, chymotrypsin and elastase, subunit III hydrolyzed p-nitrophenyl acetate at a rate similar to that of chymotrypsin but without any burst of p-nitrophenol, which indicates that the weakly functional active site of the subunit is not quite comparable to that of serine protease zymogens. Subunit III already has some of the functional characteristics of the corresponding active enzymes.
mesh_terms: D000818:Animals; D002268:Carboxypeptidases / Q000302:isolation & purification* / Q000378:metabolism; D043422:Carboxypeptidases A; D002417:Cattle; D055598:Chemical Phenomena; D002621:Chemistry; D004792:Enzyme Precursors / Q000302:isolation & purification* / Q000378:metabolism; D004950:Esterases; D005663:Furans; D006868:Hydrolysis; D008299:Maleic Anhydrides; D010179:Pancreas / Q000201:enzymology*; D010446:Peptide Fragments / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D004792:Enzyme Precursors; D005663:Furans; D008299:Maleic Anhydrides; D010446:Peptide Fragments; C007474:2,3-dimethylmaleic anhydride; D004950:Esterases; D002268:Carboxypeptidases; D043422:Carboxypeptidases A