Purification and characterization of a new metalloproteinase with elastolytic activity from the catfish pancreas.
An elastolytic enzyme was purified to homogeneity from the pancreas of the catfish Parasilurus asotus by chromatography on CM-cellulose column and affinity chromatography on (Ala)3-CH-Sepharose 4B column. The molecular weight of the enzyme was estimated to be 24 000 by SDS-polyacrylamide gel electrophoresis. The enzyme had elastolytic activity as well as activity toward Suc-(Ala)3-NA. The enzyme was inhibited by EDTA, EGTA, o-phenanthroline, dithiothreitol, and cysteine, but not by the serine proteinase inhibitors iPr2P-F and PhCH2SO2F. In addition, the enzyme was found to require Zn2+ for activity. These results indicate that the catfish enzyme belongs to the group of metalloproteinases and that it is a new type of pancreatic enzyme, unlike the pancreatic elastases which are serine proteinases.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002413:Cations, Divalent; D010450:Endopeptidases / Q000302:isolation & purification / Q000378:metabolism*; D005399:Fishes; D007700:Kinetics; D008666:Metalloendopeptidases; D008667:Metalloproteins / Q000378:metabolism*; D010179:Pancreas / Q000201:enzymology*; D010196:Pancreatic Elastase / Q000378:metabolism*; D011480:Protease Inhibitors / Q000494:pharmacology; D015032:Zinc / Q000032:analysis
D000596:Amino Acids; D002413:Cations, Divalent; D008667:Metalloproteins; D011480:Protease Inhibitors; D010450:Endopeptidases; D010196:Pancreatic Elastase; D008666:Metalloendopeptidases; D015032:Zinc