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Thromb Haemost 2001; 86(06): 1466-1474
DOI: 10.1055/s-0037-1616750
Review Article
Schattauer GmbH

Identification of Critical Residues on Thrombin Mediating Its Interaction with Fibrin

Scott W. Hall
1   Division of Hematology, Stanford University School of Medicine, Stanford, California, USA and Gilead Sciences, Foster City, California, USA
,
Craig S. Gibbs
1   Division of Hematology, Stanford University School of Medicine, Stanford, California, USA and Gilead Sciences, Foster City, California, USA
,
Lawrence L. K. Leung
1   Division of Hematology, Stanford University School of Medicine, Stanford, California, USA and Gilead Sciences, Foster City, California, USA
› Author Affiliations
Further Information

Publication History

Received 29 March 2001

Accepted after resubmission 25 July 2001

Publication Date:
12 December 2017 (online)

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Summary

Thrombin binding to fibrin may be important in localizing thrombin to the site of vascular injury. However, fibrin-bound thrombin retains its catalytic activity toward fibrinogen, and may be prothrombotic under certain conditions. A collection of 52 purified thrombin mutants was used to identify those residues mediating the thrombin-fibrin interaction. Comparison of fibrinogen clotting activity with fibrin binding activity identified twenty residues involved in fibrinogen recognition with four of these residues important in fibrin binding (Lys65, His66, Tyr71, Arg73). No mutant was identified with normal clotting activity and deficient fibrin binding, suggesting that these two properties are not readily dissociable. A DNA thrombin aptamer that binds to these residues was able to inhibit the thrombin-fibrin interaction, and displace thrombin that was already bound. Mapping of these fibrin-binding residues on thrombin revealed that they are localized within exosite I, and comprise a subset of the residues important in fibrinogen recognition.

Keywords

Thrombin - fibrin - alanine-scanning mutagenesis - clot-bound thrombin
 

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