title: Purification and characterisation of swine serum proteinase which hydrolyses epidermal inhibitory pentapeptide.
abstract: In this paper we describe the purification to molecular homogeneity of the enzyme that cleaves the synthetic epidermal mitosis-inhibiting pentapeptide (pyroGlu-Glu-Asp-Ser-Gly; EPP) from swine serum. Biochemical characterisation of the enzyme shows a glycoprotein with apparent molecular mass of 200 kDa. The Km and Kcat values for EPP hydrolysis are 0.624 mM and 694 s-1, respectively. Use of proteinase inhibitors shows the enzyme's metalloendopeptidase character. Moreover, captopril and lisinopril prevent the cleavage of EPP. The N-terminal amino-acid sequence of the purified protein corresponds to the N-terminal amino-acid sequence of swine kidney angiotensin-converting enzyme, a dipeptidyl carboxypeptidase (EC 3.4.15.1).
authors.affiliation: Department of Molecular, Cellular and Animal Biology, University of Camerino, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Bramucci
authors.forename: A
authors.initials: A
authors.lastname: Miano
authors.forename: L
authors.initials: L
authors.lastname: Quassinti
authors.forename: E
authors.initials: E
authors.lastname: Maccari
authors.forename: S
authors.initials: S
authors.lastname: Canofeni
authors.forename: D
authors.initials: D
authors.lastname: Amici
mesh_terms: D000595:Amino Acid Sequence; D000806:Angiotensin-Converting Enzyme Inhibitors / Q000494:pharmacology; D000818:Animals; D002216:Captopril / Q000494:pharmacology; D002268:Carboxypeptidases / Q000097:blood* / Q000737:chemistry; D004791:Enzyme Inhibitors / Q000494:pharmacology; D006023:Glycoproteins / Q000737:chemistry; D006131:Growth Inhibitors / Q000097:blood / Q000378:metabolism*; D007700:Kinetics; D017706:Lisinopril / Q000494:pharmacology; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000378:metabolism*; D007703:Peptidyl-Dipeptidase A / Q000378:metabolism*; D011761:Pyrrolidonecarboxylic Acid / Q000031:analogs & derivatives; D013379:Substrate Specificity; D013552:Swine
chemical_list: D000806:Angiotensin-Converting Enzyme Inhibitors; D004791:Enzyme Inhibitors; D006023:Glycoproteins; D006131:Growth Inhibitors; D009842:Oligopeptides; C050550:epidermal pentapeptide; D002216:Captopril; D017706:Lisinopril; D002268:Carboxypeptidases; D007703:Peptidyl-Dipeptidase A; D011761:Pyrrolidonecarboxylic Acid