Purification and properties of a peptidase acting on a synthetic substrate for collagenase from monkey kidney.
A peptidase cleaving a synthetic substrate for collagenase, 4-phenylazobenzyloxycarbonyl-L-Pro-L-Leu-Gly-L-Pro-D-Arg (designated as PZ-peptide) has been purified extensively (about 5200-fold) from a soluble extract of monkey kidney with a view of carrying out studies on its possible physiological role. The purified PZ-peptidase appeared essentially free of collagenase, nonspecific protease and di- and tri-peptidase activities. The properties of the purified PZ-peptidase resemble very much the granuloma enzyme. It is optimally active around pH 7.0. Its apparent Km value for PZ-peptide is 0.72 mM and V is 10.1 mumol/mg protein/min. It is reversibly inhibited by p-hydroxymercuribenzoate and HgCl2, whereas iodoactetamide does not affect the enzyme activity. N-Ethylmaleimide inhibited the enzyme partially (50%). Heavy metals like Cu-2+, Cd-2+, Ag+, Pb-2+, Ni-2+, and Zn-2+ completely inhibited the enzyme activity, while the inhibition by Co-2+ was only partial. Fe-2+ did not exert any effect on the activity. The enzyme activity was completely inhibited by EDTA and was restored almost to the original value by metal ions like Mn-2+, Mg-2+, Ca-2+ and Ba-2+. The approximate molecular weight of the purified enzyme was estimated to be 56 000.
D000818:Animals; D002413:Cations, Divalent / Q000494:pharmacology; D004151:Dipeptides; D004492:Edetic Acid / Q000494:pharmacology; D010450:Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D005033:Ethylmaleimide / Q000494:pharmacology; D005260:Female; D006902:Hydroxymercuribenzoates / Q000494:pharmacology; D007668:Kidney / Q000201:enzymology*; D007700:Kinetics; D008251:Macaca; D008297:Male; D008628:Mercury / Q000494:pharmacology; D003012:Microbial Collagenase / Q000032:analysis; D008970:Molecular Weight; D009842:Oligopeptides; D010455:Peptides
D002413:Cations, Divalent; D004151:Dipeptides; D006902:Hydroxymercuribenzoates; D009842:Oligopeptides; D010455:Peptides; D004492:Edetic Acid; D010450:Endopeptidases; D003012:Microbial Collagenase; D008628:Mercury; D005033:Ethylmaleimide