Action of crotalase, an enzyme with thrombin-like and kallikrein-like specificities, on tripeptide nitroanilide derivatives.
Since crotalase has thrombin-like and kallikrein-like functional and structural properties, we compared the actions of crotalase, thrombin and plasma kallikrein on 13 tripeptide nitroanilide substrates. Initial rates of hydrolysis were determined at 27 degrees C, pH 8.3, and used to construct Lineweaver-Burk plots from which Km and Vmax were determined. The ratio of kcat/Km was taken as a measure of enzymatic specificity. Crotalase yielded kcat/Km values for the following nitroanilide substrates in descending order of magnitude: H-D-NLeu-CHA-Arg, H-D-Pro-HHT-Arg, Tos-Gly-Pro-Arg, H-D-PhGly-Phe-Arg, Cbo-Glu(BuO)-Gly-Arg, H-D-But-CHA-Lys, H-D-CHG-But-Arg, H-D-NLeu-HHT-Lys, H-D-HHT-Ala-Arg, Bz-Pro-Phe-Arg, Tos-Gly-Pro-Lys, MeS-Leu-Gly-Arg, MeO-CO-CHG-Gly-Arg. This pattern of specificity correlated only roughly with those of thrombin and kallikrein.
D002863:Chromogenic Compounds; D003435:Crotalid Venoms; D010450:Endopeptidases / Q000378:metabolism*; D005170:Factor X / Q000032:analysis; D015951:Factor Xa; D005341:Fibrinolysin / Q000032:analysis; D006801:Humans; D006868:Hydrolysis; D007610:Kallikreins / Q000097:blood; D009842:Oligopeptides; D012697:Serine Endopeptidases; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D013917:Thrombin / Q000032:analysis; D014568:Urokinase-Type Plasminogen Activator / Q000032:analysis
D002863:Chromogenic Compounds; D003435:Crotalid Venoms; D009842:Oligopeptides; D005170:Factor X; D010450:Endopeptidases; D007610:Kallikreins; D012697:Serine Endopeptidases; D013917:Thrombin; D015951:Factor Xa; D005341:Fibrinolysin; D014568:Urokinase-Type Plasminogen Activator; C019948:crotalase