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Recombinant hirudin: kinetic mechanism for the inhibition of human thrombin

Stone, Stuart R. ; Hofsteenge, Jan

In: Protein Engineering, Design and Selection, 1991, vol. 4, no. 3, p. 295-300

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    Titre
    • Recombinant hirudin: kinetic mechanism for the inhibition of human thrombin
    Auteur
    • Stone, Stuart R.. Friedrich Miescher-Institut, PO Box 2543. CH-4002 Basel, Switzerland
    • Hofsteenge, Jan. Friedrich Miescher-Institut, PO Box 2543. CH-4002 Basel, Switzerland
    Type de document
    • Postprint
    Langue
    • Inglese
    Publié dans
    • Protein Engineering, Design and Selection, 1991, vol. 4, no. 3, p. 295-300. Oxford University Press
    Autre version électronique
    Identifiant OAI-PMH
    • oai:doc.rero.ch:290650
    Summary
    Recombinant hirudin variant-2(Lys47 ), was found to be a competitive inhibitor of human α-thrombin with respect to peptidyl p-Miitroanilide substrates. These results contrast with those of Degryse and coworkers that suggest that recombinant hirudin variant-2(Lys47) inhibited thrombin by a non-competitive mechanism [Degryse et al. (1989) Protein Engng, 2, 459-465], γ-Thrombin, which can arise from α-thrombin by autolysis, was shown to have an affinity for recombinant hirudin variant-2(Lys47) that was four orders of magnitude lower than that of α-thrombin. It was demonstrated that the apparent noncompetitive mechanism observed previously was probably caused by a contamination of the thrombin preparation by γ-thrombin. Comparison of the inhibition of α-thrombin by recombinant hirudins variant-2(Lys47) and variant-1, which differ from one another in eight out of 65 amino acids, indicated that the two variants have essentially the same kinetic parameters