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Thromb Haemost 1977; 38(04): 0776-0792
DOI: 10.1055/s-0038-1651897
Original Article
Schattauer GmbH

Serine Protease Specificity for Peptide Chromogenic Substrates

L. E Mattler
1   Lilly Laboratory for Clinical Research and Department of Medicine, Indiana University School of Medicine, Indianapolis, Indiana 46202, U.S.A.
,
N. U Bang
1   Lilly Laboratory for Clinical Research and Department of Medicine, Indiana University School of Medicine, Indianapolis, Indiana 46202, U.S.A.
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Publication History

Publication Date:
04 July 2018 (online)

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Summary

Rates of hydrolysis of the newly developed peptide chromogenic substrates S-2160 (N-Bz-Phe-Val-Arg-pNA, HCl), S-2238 (H-D-Phe-Pip-Arg-pNA, 2HC1), S-2222 (N-Bz-Ile-Glu-Gly-Arg-pNA, HC1), and S-2251 (H-D-Val-Leu-Lys-pNA, 2HC1) from AB Kabi Peptide Research and Chromozym TH (Z-Gly-Pro-Arg-pNA, HC1) from Pentapharm Limited were tested against highly purified preparations of human plasmin, bovine trypsin, human alpha thrombin, and bovine factor Xa.

S-2160, S-2238, and Chromozym TH are sensitive to thrombin, Chromozym TH and S-2238 exhibiting a substantially greater sensitivity than S-2160. All 3 substrates are insensitive to factor Xa but hydrolyzed to varying degrees by plasmin and trypsin. In contrast, S-2222 is sensitive to Xa and insensitive to thrombin. S-2251 is relatively plasmin-specific, being resistant to the clotting enzymes thrombin and Xa. S-2251 exhibits even greater sensitivity to the SK-plasmin complex than to plasmin. In addition, the substrate Chromozym PK (N-Bz-Pro-Phe-Arg-pNA, HC1) was evaluated and found to be relatively specific for plasma kallikrein. Assays for antithrombin III and heparin using S-2222 as the substrate and factor Xa as the enzyme, plasma plasminogen and plasmin inhibitors using S-2251 as the substrate, and plasma prekallikrein and kallikrein inhibitors using Chromozym PK as the substrate have been developed. Synthetic peptides mimicking amino acid sequences adjacent to proteolytic activation cleavage of plasma serine protease precursors appear to be sensitive and relatively specific tools applicable to kinetical and clinical studies of these enzymes and their inhibitors.

 

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