Role of hydrophobic mutations on the binding affinity and stability of blood coagulation factor VIIIa: a computational molecular dynamics and free-energy analysis.
Role of hydrophobic mutations on the binding affinity and stability of blood coagulation factor VIIIa: a computational molecular dynamics and free-energy analysis.
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DOI：
10.1016/j.bbrc.2014.06.043
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发表时间：
2014-07-18
影响因子：
3.1
通讯作者：
Venkateswarlu, Divi
中科院分区：
文献类型：
--
作者：
Venkateswarlu, Divi
因子VIIIa是A1、A2和A3-C1-C2结构域之间的非共价结合的异源三聚体，并且在因子X酶原的蛋白水解活化期间是因子IXa酶的必需辅因子。A2和界面A1/A3结构域之间相对弱的相互作用抑制了血浆中FVIIIa的功能稳定性并导致快速降解。采用分子动力学模拟方法研究了三个带电残基（Asp 519、Glu 665和Asp 666）对几个疏水残基的突变效应。通过MM-PBSA和MM-GBSA方法分析结合自由能表明，Asp 519和Glu 665残基突变为瓦尔或Ala增强了A2结构域结合亲和力，与实验位点特异性诱变数据一致。Asp 666突变为瓦尔、Tyr、Met和Phe在所研究的8个疏水突变体中显示出A2结构域结合的最大改善。我们的研究表明，在A2结构域的掩埋表面区域中的疏水相互作用的富集在提高FVIIIa的整体稳定性中起着至关重要的作用。
Factor VIIIa is a non-covalently bound hetero-trimer among A1, A2 and A3-C1-C2 domains and an essential co-factor for factor IXa enzyme during proteolytic activation of factor X zymogen. The relatively weak interactions between A2 and the interface A1/A3 domains dampen the functional stability of FVIIIa in plasma and results in rapid degradation. We studied the mutational effect of three charged residues (Asp519, Glu665 and Asp666) to several hydrophobic residues by molecular dynamics simulations. Analysis of the binding free energy by MM-PBSA and MM-GBSA methods shows that the mutation of Asp519 and Glu665 residues to either Val or Ala enhance the A2 domain binding affinity in agreement with the experimental site-specific mutagenesis data. Mutation of Asp666 to Val, Tyr, Met and Phe showed largest improvement in the A2-domain binding among the eight hydrophobic mutants studied. Our studies suggest that the enrichment of hydrophobic interactions in the buried surface regions of A2 domain plays crucial role in improving the overall stability of FVIIIa.
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