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  • New fluorogenic substrates for alpha-thrombin, factor Xa, kallikreins, and urokinase.

New fluorogenic substrates for alpha-thrombin, factor Xa, kallikreins, and urokinase.

Journal of biochemistry (1977-11-01)
T Morita, H Kato, S Iwanaga, K Takada, T Kimura
PMID591514
ABSTRACT

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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Pro-Phe-Arg-7-amido-4-methylcoumarin, ≥95%
Sigma-Aldrich
Z-Phe-Arg-7-amido-4-methylcoumarin, Hydrochloride, Substrate for fluorogenic assay of plasma and glandular kallikreins.
Sigma-Aldrich
Boc-Val-Pro-Arg-7-amido-4-methylcoumarin hydrochloride
Sigma-Aldrich
Z-Phe-Arg 7-amido-4-methylcoumarin hydrochloride, kallikrein substrate
Sigma-Aldrich
Boc-Ile-Glu-Gly-Arg-7-amido-4-methylcoumarin acetate salt, ≥97% (HPLC)