Mapping the interaction between factor VIII and von Willebrand factor by electron microscopy and mass spectrometry
Mapping the interaction between factor VIII and von Willebrand factor by electron microscopy and mass spectrometry
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DOI：
10.1182/blood-2015-04-641688
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发表时间：
2015-08-20
期刊：
影响因子：
20.3
通讯作者：
Walz, Thomas
中科院分区：
文献类型：
--
作者：
Chiu, Po-Lin;Bou-Assaf, George M.;Walz, Thomas
与血管性血友病因子(VWF)的D‘D3结构域结合可稳定循环中的因子VIII(FVIII)，并将其维持在足以防止自发性出血的水平。我们使用负染色电子显微镜(EM)观察了FVIII与D‘D3结构域的二聚体和单体形式的络合物。EM平均结果表明，FVIII主要通过其C1域与D‘D3结构域相互作用，C2结构域提供第二个连接位点。氢-氚交换质谱仪证实了C1结构域在D‘D3结合中的重要性，并暗示了FVIII上额外的表面区在相互作用中。综上所述，我们的结果证实了C1域是VWF与FVIII的主要结合部位，重申了A3酸性肽在VWF结合中的重要性，并表明A3和C2结构域在这种相互作用中起辅助作用。
Association with the D'D3 domain of von Willebrand factor (VWF) stabilizes factor VIII (FVIII) in the circulation and maintains it at a level sufficient to prevent spontaneous bleeding. We used negative-stain electron microscopy (EM) to visualize complexes of FVIII with dimeric and monomeric forms of the D'D3 domain. The EM averages show that FVIII interacts with the D'D3 domain primarily through its C1 domain, with the C2 domain providing a secondary attachment site. Hydrogen-deuterium exchange mass spectrometry corroborated the importance of the C1 domain in D'D3 binding and implicates additional surface regions on FVIII in the interaction. Together, our results establish that the C1 domain is the major binding site on FVIII for VWF, reiterate the importance of the a3 acidic peptide in VWF binding, and suggest that the A3 and C2 domains play ancillary roles in this interaction.