Backbone resonance assignments of the A2 domain of mouse von Willebrand factor
Backbone resonance assignments of the A2 domain of mouse von Willebrand factor
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DOI：
10.1007/s12104-021-10041-8
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发表时间：
2021-07-20
影响因子：
0.9
通讯作者：
Sugase，Kenji
中科院分区：
文献类型：
--
作者：
Morimoto，Daichi;Osugi，Masanori;Sugase，Kenji
血管性血友病因子 (vWF) 是一种血浆粘附蛋白，对于血管损伤后正常止血中的血小板粘附非常重要。尽管响应止血刺激，血小板和（亚）内皮细胞的储存颗粒会释放大的 vWF 多聚体，但为了正常的生理功能，需要将 vWF 多聚体裂解成更小的多聚体形式。血浆金属蛋白酶 ADAMTS13 特异性裂解位于 vWF (vWF-A2) A2 结构域中部的肽键，但裂解位点埋藏在 vWF 结构内部，在没有升高的流动剪切应力的情况下很难进入。另一方面，在存在高血管剪切应力的情况下，vWF-A2 的结构应该会展开，从而易于被 ADAMTS13 进行蛋白水解。然而，vWF-A2 剪切引起的结构变化背后的原子级机制仍不清楚，并且迄今为止还没有可用的解决方案 NMR 信息。在本研究中，我们展示了小鼠 vWF-A2 的主链 1H、13C 和 15N 共振分配；还提供了 13 Cβ 的侧链分配。基于指定化学位移的二级结构倾向分析表明，小鼠vWF-A2在溶液中形成与先前确定的人vWF-A2晶体结构相似的二级结构。获得的 NMR 分配数据将有助于溶液中 vWF-A2 剪切诱导展开的原子级表征。
von Willebrand factor (vWF) is an adhesive plasma protein that is important for platelet adhesion in normal hemostasis in response to vascular injury. Although large vWF multimers are released from storage granules of platelets and (sub-)endothelial cells in response to hemostatic stimuli, for normal physiological function, vWF multimers are required to be cleaved into smaller multimeric forms. The plasma metalloproteinase ADAMTS13 specifically cleaves the peptide bond located in the middle of the A2 domain of vWF (vWF-A2), but the cleavage site is buried inside the structure of vWF and is difficult to access in the absence of elevated flow shear stress. On the other hand, in the presence of high vascular shear stress, the structure of vWF-A2 is supposed to be unfolded, thereby becoming accessible for proteolysis by ADAMTS13. However, the atomic-level mechanism underlying shear-induced structural changes of vWF-A2 remains unclear and to date no solution NMR information is available. In this study, we present the backbone1H,13C, and15N resonance assignments of mouse vWF-A2; side chain assignments of13Cβare also provided. Secondary structure propensity analysis based on the assigned chemical shifts showed that mouse vWF-A2 forms similar secondary structures in solution to the previously determined crystal structure of human vWF-A2. The obtained NMR assignment data will contribute to an atomic-level characterization of shear-induced unfolding of vWF-A2 in solution.