Fibrinolysis with des-kringle derivatives of plasmin and its modulation by plasma protease inhibitors

Erzsébet Komorowicz, Krasimir Kolev, Raymund Machovich

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26 Citations (Scopus)

Abstract

Quantitative characterization of the interaction of des-kringle1-5- plasmin (microplasmin) with fibrin(ogen) and plasma protease inhibitors may serve as a tool for further evaluation of the role of kringle domains in the regulation of fibrinolysis. Comparison of fibrin(ogen) degradation products yielded by plasmin, miniplasmin (des-kringle1-4-plasmin), microplasmin, and trypsin on SDS gel electrophoresis indicates that the differences in the enzyme structure result in different rates of product formation, whereas the products of the four proteases are very similar in molecular weight. Kinetic parameters show that plasmin is the most efficient enzyme in fibrinogen degradation, and the k(cat)/K(M) ratio decreases in parallel with the loss of the kringle domains. The catalytic sites of the four proteases have similar affinities for fibrin (K(M) values between 0.12 and 0.21 μM). Trypsin has the highest catalytic constant for fibrin digestion (k(cat) = 0.47 s-1), and among plasmins with different kringle structures, the loss of kringle5 results in a markedly lower catalytic rate constant (k(cat) = 0.0076 s-1 for microplasmin vs 0.048 s-1 for miniplasmin and 0.064 s-1 for plasmin). In addition, microplasmin is inactivated by plasmin inhibitor (k'' = 3.9 x 105 M-1 s-1) and antithrombin (k'' = 1.4 x 103 M-1 s-1) and the rate of inactivation decreases in the presence of fibrin(ogen). Heparin (250 nM) accelerates the inactivation of microplasmin by antithrombin (k'' = 10.5 x 103 M-1 s-1), whereas that by plasmin inhibitor is not affected (k'' = 4.2 x 105 M-1 s-1).

Original languageEnglish
Pages (from-to)9112-9118
Number of pages7
JournalBiochemistry
Volume37
Issue number25
DOIs
Publication statusPublished - Jun 23 1998

ASJC Scopus subject areas

  • Biochemistry

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