title: Preparation and characterization of novel substrates of insulin proteinase (EC 3.4.99.45).
abstract: The specificity of insulin proteinase (EC 3.4.99.45) has been difficult to categorize using only its natural substrates. By exploiting the fact that two substrates competing for the same enzyme inhibit one another, we have found some new substrates of the insulin proteinase from porcine muscle. Two of these substrates, a tryptic fragment of BSA and a fragment of cytochrome c, have been shown to be cleaved at a single site. The albumin fragment, as well as another fragment of cytochrome c., have susceptibilities (Vmax/Km) comparable with that of insulin. In a second aspect of the study, the porcine-muscle enzyme was shown to be related to other members of its superfamily in that it was immunoprecipitated by a monoclonal antibody raised against the insulin-degrading enzyme from human red blood cells and has the same cleavage sites on insulin as has the rat skeletal-muscle insulin proteinase. We note, however, a possible discrepancy between our results and those of another group regarding the subunit size (110 kDa) of the immunoprecipitated material.
authors.affiliation: Département de Biochimie Médicale, C.M.U., Geneva, Switzerland.
authors.forename: R C
authors.initials: RC
authors.lastname: Werlen
authors.forename: R E
authors.initials: RE
authors.lastname: Offord
authors.forename: K
authors.initials: K
authors.lastname: Rose
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D003574:Cytochrome c Group / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D007339:Insulysin / Q000276:immunology / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009132:Muscles / Q000201:enzymology*; D010446:Peptide Fragments / Q000378:metabolism; D011233:Precipitin Tests; D011863:Radioimmunoassay; D051381:Rats; D012710:Serum Albumin, Bovine / Q000737:chemistry / Q000378:metabolism*; D013379:Substrate Specificity; D013552:Swine
chemical_list: D000911:Antibodies, Monoclonal; D003574:Cytochrome c Group; D010446:Peptide Fragments; D012710:Serum Albumin, Bovine; D007339:Insulysin