Antithrombin properties of C‐terminus of hirudin using synthetic unsulfated N α‐acetyl‐hirudin45–65

FEBS Letters

Mao, Simon J.T.¹  Krstenansky, John L.¹ 

[1] Merrel Dow Research Institute, 2110 E. Galbraith Rd, Cincinnati, OH 45215, USA

关键词: Thrombin;    Hirudin;    Thrombin inhibitor;    Fibrinogen clot;    (Leech);    Boc;    t-butoxycarbonyl;    2-BrZ;    2-bromobenzyloxycarbonyl;    Bzl;    benzyl;    Chx;    cyclohexyl;    2;    6-diClZ;    2;    6-dichlorobenzyloxycarbonyl;    DCC;    N;    N'-dicyclohexylcarbodiimide;    FAB-MS;    fast atom bombardment mass spectrometry;    HOBT;    1-hydroxybenzo-triazole;    HPLC;    high-performance liquid chromatography;    PAM;    phenylacetamidomethyl;    Pip;    piperidine-2-carboxylate;    TLC;    thin layer chromatography;   

DOI  :  10.1016/0014-5793(87)81264-4

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Unsulfated N ^α-acetyl-hirudin_45–65 (MDL 27 589), which corresponds to the C-terminus of hirudin_1–65, was synthesized by solid-phase methods. The synthetic peptide was able to inhibit fibrin formation and the release of fibrinopeptide A from fibrinogen by thrombin. The catalytic site of thrombin was not perturbed by the synthetic peptide as H-D-Phe-Pip-Arg-pNA hydrolysis (amidase activity) was not affected. The binding of synthetic peptide and thrombin was assessed by isolation of the complex on gel-filtration chromatography. A single binding site with a binding affinity (K _a) of approx. 1.0 × 10⁵ M⁻¹ was observed for thrombin-hirudin_45–65 interaction. The data suggest that the C-terminal residues 45–65 of hirudin contain a binding domain which recognizes thrombin and yet does not bind to the catalytic site of the enzyme.

【 授权许可】

Unknown