Identification of Critical Residues on Thrombin Mediating Its Interaction with Fibrin

Scott W. Hall; Craig S. Gibbs; Lawrence L. K. Leung

doi:10.1055/s-0037-1616750

Thrombosis and Haemostasis, Table of Contents

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

Authors

Scott W. Hall

¹Division of Hematology, Stanford University School of Medicine, Stanford, California, USA and Gilead Sciences, Foster City, California, USA
Craig S. Gibbs

¹Division of Hematology, Stanford University School of Medicine, Stanford, California, USA and Gilead Sciences, Foster City, California, USA
Lawrence L. K. Leung

¹Division of Hematology, Stanford University School of Medicine, Stanford, California, USA and Gilead Sciences, Foster City, California, USA

Abstract

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

Full Text

References

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