Purification to homogeneity and properties of two D-alanine carboxypeptidases I From Escherichia coli.
Three homogeneous preparations of D-alanine carboxypeptidases I have been obtained from Escherichia coli strain H2143, termed enzymes IA, IB, and IC. Enzyme IA purified from the membrane after extraction with Triton X-100 appeared on sodium dodecyl sulfate gel electrophoresis to be a polypeptide doublet whose monomer molecular weights were about 32,000 and 34,000. In addition to D-alanine carboxypeptidase activity, it catalyzed a transpeptidase reaction with several substrates, bound [14C]penicillin G, had a weak penicillinase activity, but was devoid of endopeptidase activity. Enzyme IB obtained from the membrane after LiCl extraction and enzyme IC obtained from the supernatant solution were either identical or extremely similar. They were composed of a single polypeptide whose monomer molecular weight was about 41,000. In addition to carboxypeptidase activity, they catalyzed an endopeptidase reaction, had weak penicillinase activity, and had very poor transpeptidase activity, but did not bind [14C]penicillin G. Some data relating to the mechanism of catalysis by these enzymes are described. Their possible physiological role is discussed.
D001665:Binding Sites; D002268:Carboxypeptidases / Q000302:isolation & purification* / Q000378:metabolism; D002413:Cations, Divalent; D002729:Chloromercuribenzoates / Q000494:pharmacology; D004492:Edetic Acid / Q000494:pharmacology; D004926:Escherichia coli / Q000201:enzymology*; D006863:Hydrogen-Ion Concentration; D007527:Isoenzymes / Q000302:isolation & purification* / Q000378:metabolism; D007700:Kinetics; D010400:Penicillin G / Q000378:metabolism; D010405:Penicillinase / Q000378:metabolism; D010458:Peptidyl Transferases / Q000378:metabolism; D011485:Protein Binding; D011955:Receptors, Drug
D002413:Cations, Divalent; D002729:Chloromercuribenzoates; D007527:Isoenzymes; D011955:Receptors, Drug; D004492:Edetic Acid; D010458:Peptidyl Transferases; D002268:Carboxypeptidases; D010405:Penicillinase; D010400:Penicillin G