Purification and characterisation of swine serum proteinase which hydrolyses epidermal inhibitory pentapeptide.
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).
Bramucci
M
M
Department of Molecular, Cellular and Animal Biology, University of Camerino, Italy.
Miano
A
A
Quassinti
L
L
Maccari
E
E
Canofeni
S
S
Amici
D
D
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
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