Use of Protein Chemistry and Molecular Biology to Determine Interaction Areas Between Proteases and their Inhibitors: The Thrombin-Hirudin Interaction as an Example

  • Stuart R. Stone
  • Stanley Dennis
  • Andrew Wallace
  • Jan Hofsteenge
Part of the NATO ASI Series book series (NSSA, volume 191)


Thrombin is a serine protease that exhibits the same primary specificity as trypsin, i.e., it cleaves peptide bonds on the C-terminal side of basic amino acids (preferably arginine) Thrombin, however, exhibits a narrower specificity than trypsin with respect to the peptide bonds that it will cleave. This narrower specificity of thrombin is partly due to the presence on thrombin of secondary binding sites and this article will concentrate on the contribution of these sites to the formation of the complex between thrombin and the inhibitor hirudin. After a brief introduction on thrombin, hirudin and the kinetics of the formation of their complex, studies aimed at identifying secondary binding sites for hirudin on thrombin will be considered. These studies have used mainly the techniques of protein chemistry. The last part of the article will consider the use of site-directed mutagenesis to elucidate areas of hirudin important for its inhibitory activity.


Serine Lysine Arginine Glutamine Trypsin 


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Copyright information

© Plenum Press, New York 1990

Authors and Affiliations

  • Stuart R. Stone
    • 1
  • Stanley Dennis
    • 1
  • Andrew Wallace
    • 1
  • Jan Hofsteenge
    • 1
  1. 1.Friedrich Miescher-InstitutBaselSwitzerland

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