title: Comparison of the enzymatic and biochemical properties of human insulin-degrading enzyme and Escherichia coli protease III.
abstract: The enzymatic and biochemical properties of human insulin-degrading enzyme and Escherichia coli protease III have been compared. Both enzymes were found to degrade insulin in such a way that its receptor binding activity was rapidly lost but its precipitability in trichloracetic acid was only slightly decreased. Both enzymes were also found to be inhibited by chelating agents. The bacterial enzyme, which could be purified in large amounts, was found to contain 0.6 mol of zinc per mol of enzyme but no detectable manganese. The mammalian enzyme but not the bacterial one was inhibited by a sulfhydryl alkylating agent. The two enzymes also differed in substrate specificity. The mammalian enzyme degraded insulin much better than insulin-like growth factor II, whereas the bacterial enzyme degraded them equally. The mammalian enzyme could be labeled by cross-linking to insulin = bombyxin II much greater than insulin-like growth factor I and II much greater than relaxin, while the bacterial enzyme was labeled by insulin-like growth factor II greater than insulin = insulin-like growth factor I much greater than relaxin much greater than bombyxin. Finally, sucrose gradient centrifugation and cross-linking studies both in vitro and in vivo indicated that active human enzyme partially existed as a homo- or heterodimer, whereas the bacterial enzyme was active as a monomer.
authors.affiliation: Department of Pharmacology, Stanford University School of Medicine, California 94305-5332.
authors.forename: L
authors.initials: L
authors.lastname: Ding
authors.forename: A B
authors.initials: AB
authors.lastname: Becker
authors.forename: A
authors.initials: A
authors.lastname: Suzuki
authors.forename: R A
authors.initials: RA
authors.lastname: Roth
mesh_terms: D000477:Alkylating Agents / Q000494:pharmacology; D000818:Animals; D000911:Antibodies, Monoclonal; D001345:Autoradiography; D015153:Blotting, Western; D006224:Cricetinae; D003412:Cricetulus; D003432:Cross-Linking Reagents; D004591:Electrophoresis, Polyacrylamide Gel; D010450:Endopeptidases / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D004912:Erythrocytes / Q000201:enzymology; D004926:Escherichia coli / Q000201:enzymology*; D005260:Female; D006801:Humans; D007339:Insulysin / Q000037:antagonists & inhibitors / Q000378:metabolism*; D008666:Metalloendopeptidases; D051379:Mice; D008807:Mice, Inbred BALB C; D011480:Protease Inhibitors / Q000494:pharmacology; D012065:Relaxin / Q000378:metabolism; D013002:Somatomedins / Q000378:metabolism; D013379:Substrate Specificity; D013438:Sulfhydryl Compounds / Q000494:pharmacology
chemical_list: D000477:Alkylating Agents; D000911:Antibodies, Monoclonal; D003432:Cross-Linking Reagents; D011480:Protease Inhibitors; D013002:Somatomedins; D013438:Sulfhydryl Compounds; D012065:Relaxin; D010450:Endopeptidases; D008666:Metalloendopeptidases; C046583:pitrilysin; D007339:Insulysin