Interaction of site specific hirudin variants with α‐thrombin

FEBS Letters

Baici, Antonio¹  Dodt, Johannes²  Köhler, Stefanie² 

[1] Forschungslabor der Rheumaklinik, Universitätsspital, Gloriastr. 25, CH-8091 Zürich, Switzerland;Institut für Biochemie der Technischen Hochschule Darmstadt, Petersenstr. 22, D-6100 Darmstadt, FRG

关键词: Hirudin;    Hirudin variant;    Site-directed mutagenesis;    Kinetics;    α-Thrombin;    Putative reactive site;    HPLC;    high-performance liquid chromatography;    IPTG;    isopropyl-β-D-thiogalactopyranoside;    RP;    reversed-phase;    Tos;    4-toluoslsulfonyl;   

DOI  :  10.1016/0014-5793(88)80803-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The kinetics of complex formation between recombinant hirudin or recombinant hirudin mutants with thrombin were analyzed. In order to elucidate the inhibitor's reactive site peptide bond predetermined amino acid substitutions were introduced at positions of basic amino acid residues by means of site-directed mutagenesis of a hirudin gene. In comparison to recombinant hirudin (K _i = 19 pM) only those mutant inhibitors which were modified at amino acid position Lys⁴⁷ showed a higher K _i value for their complexes with thrombin. The observed effects are mainly due to increased k _off rate constants.

【 授权许可】

Unknown