Interactions of hirudin-based inhibitor with thrombin: critical role of the IleH59 side chain of the inhibitor.


:Hirudin is the most potent and specific thrombin inhibitor from medicinal leech with a Ki value of 2.2 x 10(-14) M. It consists of an active site inhibitor segment, hirudin1-48, a fibrinogen-recognition exosite inhibitor segment, hirudin55-65, and a linker, hirudin49-54, connecting these inhibitor segments. The role of the side chain of the hirudin 59th residue, Ile, is studied by using a series of synthetic bivalent thrombin inhibitors, which mimic the binding mode of hirudin. The synthetic inhibitors based on the hirudin sequence have a general sequence of Ac-(D-Phe)-Pro-Arg-Pro-(4-aminobutyric acid)-(7-amino-heptanoic acid)-Asp-Phe-Glu-Glu-Xaa-Pro-Glu-Glu-Tyr-Leu-Gln-OH, in which the 59th residue, Xaa, is substituted by various natural and unnatural L-amino acids. For example, substitution of IleH59 by Val, which is equivalent to removing the delta-methyl group of IleH59, reduces the affinity of the inhibitor 5.7-fold (delta delta G0 = 1.0 kcal/mol) to a Ki value of 4.7 nM compared to that (Ki = 0.82 nM) of the corresponding inhibitor with IleH59. Removal of the entire side chain of IleH59, i.e., a substitution of IleH59 by Gly, reduces the affinity of the inhibitor 6300-fold, revealing the critical role of the IleH59 side chain in the inhibitor binding. Theoretical free energy calculation successfully reproduces the binding free energy of most of the analogs. It suggests that intra- and intermolecular van der Waals interactions of delta-CH3, gamma-CH3, and gamma-CH2 of IleH59 play the major role in the binding affinity.(ABSTRACT TRUNCATED AT 250 WORDS)






Tsuda Y,Szewczuk Z,Wang J,Yue SY,Purisima E,Konishi Y




Has Abstract


1995-07-11 00:00:00












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    pub_type: 杂志文章


    authors: Rand-Weaver M,Noso T,Muramoto K,Kawauchi H

    更新日期:1991-02-12 00:00:00

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    pub_type: 杂志文章


    authors: Faller LD

    更新日期:1989-08-08 00:00:00

  • Tyr51: Key Determinant of the Low Thermostability of the Colwellia psychrerythraea Cold-Shock Protein.

    abstract::Cold-shock proteins (Csps) are expressed at lower-than-optimum temperatures, and they function as RNA chaperones; however, no structural studies on psychrophilic Csps have been reported. Here, we aimed to investigate the structure and dynamics of the Csp of psychrophile Colwellia psychrerythraea 34H, ( Cp-Csp). Althou...


    pub_type: 杂志文章


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    更新日期:2018-07-03 00:00:00

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    pub_type: 杂志文章


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    更新日期:2014-02-18 00:00:00

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    更新日期:2015-01-20 00:00:00

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    更新日期:1993-09-14 00:00:00

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    更新日期:2019-09-10 00:00:00

  • Type III effector NleH2 from Escherichia coli O157:H7 str. Sakai features an atypical protein kinase domain.

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    pub_type: 杂志文章


    authors: Hayes GR,Williams AM,Lucas JJ,Enns CA

    更新日期:1997-04-29 00:00:00

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    pub_type: 杂志文章


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    更新日期:1991-08-27 00:00:00

  • Characterization of the phosphoserine of pepsinogen using 31P nuclear magnetic resonance: corroboration of X-ray crystallographic results.

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    pub_type: 杂志文章


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    更新日期:1986-10-21 00:00:00

  • Redesign of the interior hydrophilic region of mitochondrial cytochrome c by site-directed mutagenesis.

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    pub_type: 杂志文章


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    pub_type: 杂志文章


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    更新日期:1975-08-26 00:00:00

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    pub_type: 杂志文章


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    pub_type: 杂志文章


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