Purification and properties of a collagen peptidase (PZ-peptidase) from rabbit serum.
A peptidase cleaving a synthetic substrate for collagen peptidases, 4-phenylazobenzyloxcarbonyl-L-Pro-L-Leu-Gly-L-pro-D-Arg (designated as PZ-peptide) has been purified 1200-fold from rabbit serum and characterized. The enzyme preparation is free of collagenase and unspecific proteinase activity. The natural substrates are denatured collagen and collagen peptides. The peptidase has a molecular weight of 124 000 and an isoelectric point at pH 5.1. The pH dependence curve exhibits two maxima, one at pH 7.1 and the other at pH 7.9. The enzymic reaction is completely inhibited by Zn2+ and to a slower degree by Hg2+, Mn2+ and p-hydroxymercuribenzoate. It is not affected by EDTA and KCN but totally blocked by o-phenanthroline. Phenylmethylsulfonylfluoride is completely inhibitory and points to a serine residue in the active site.
D000818:Animals; D000838:Anions / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D003012:Microbial Collagenase / Q000097:blood*; D008970:Molecular Weight; D009842:Oligopeptides / Q000378:metabolism; D011480:Protease Inhibitors; D011817:Rabbits; D013379:Substrate Specificity
D000838:Anions; D009842:Oligopeptides; D011480:Protease Inhibitors; D003012:Microbial Collagenase