New fluorogenic substrates for alpha-thrombin, factor Xa, kallikreins, and urokinase.
Twenty peptide-4-methylcoumarin amides (MCA) were newly synthesized and tested as possible substrates for alpha-thrombin, factor Xa, kallikreins, urokinase, and plasmin. These fluorogenic peptides contained arginine-MCA as the carboxyl-terminus. Release of 7-amino-4-methylcoumarin was determined fluorometrically. Of these peptides, the following were found to be specific substrates for individual enzymes: Boc-Val-Pro-Arg-MCA for alpha-thrombin, Boc-Ile-Glu-Gly-Arg-MCA, and Boc-Ser-Gly-Arg-MCA for factor Xa, Z-Phe-Arg-MCA for plasma kallikrein, Pro-Phe-Arg-MCA for pancreatic and urinary kallikreins, and glutaryl-Gly-Arg-MCA for urokinase. Moreover, these peptide-MCA substrates were resistant to plasmin.
D000818:Animals; D003374:Coumarins; D010450:Endopeptidases / Q000378:metabolism*; D005170:Factor X / Q000378:metabolism*; D007610:Kallikreins / Q000378:metabolism*; D010179:Pancreas / Q000201:enzymology; D013329:Structure-Activity Relationship; D013379:Substrate Specificity; D013917:Thrombin / Q000378:metabolism*; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism*
D003374:Coumarins; D005170:Factor X; D010450:Endopeptidases; D007610:Kallikreins; D013917:Thrombin; D014568:Urokinase-Type Plasminogen Activator