Structural basis for the cleavage of VWF by ADAMTS-13 under shear forces

ADAMTS-13 在剪切力下裂解 VWF 的结构基础

• 批准号: 20790030
• 负责人: NISHIDA Noritaka
• 金额: $ 2.75万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Young Scientists (B)
• 财政年份: 2008
• 资助国家: 日本
• 起止时间: 2008 至 2009
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20790030/
• 关键词: 構造生物学; NMR; 血小板凝集; VWF; ADAMTS-13

Von Willebrand factor (VWF) mediates platelet adhesion under high shear stress. The activity of VWF is controlled by the ADAMTS13, which cuts the VWF in the force dependent manner. However, the structural basis as to how VWF exposes the cryptic site in the A2 domain for ADAMTS13 is unknown. In this study, I made a recombinant VWF A2 domain, using Pichia Pastorsis. Backbone resonance assignment of A2 domain was established for >80% signals. Based on the chemical shift change induced by low concentration of urea, the a3 and a5 helices and a4less loop, which is proximal to the scissile bond, undergoes a conformational change that would exposes the cleavage site for ADAMTS13.

血管性血友病因子（VWF）介导高剪切应力下血小板粘附。VWF的活性由ADAMTS13控制，ADAMTS13以力依赖的方式切割VWF。然而，关于VWF如何暴露ADAMTS13 A2结构域的神秘位点的结构基础尚不清楚。在本研究中，我利用毕赤酵母构建了重组VWF A2结构域。对> ~ 80%的信号建立A2域的骨干共振分配。基于低浓度尿素引起的化学位移变化，位于可剪键附近的a3和a5螺旋和a4less环发生构象变化，暴露出ADAMTS13的裂解位点。

项目成果

Structure of an integrin with an αl domain, complement receptor type 4

具有 α1 结构域的整联蛋白的结构，补体受体 4 型

• 发表时间: 2010
• 期刊: The EMBO Journal 29
• 作者: Xie C;Zhu J;Chen X;Mi LZ;Nishida N;Springer TA;Can Xie
• 通讯作者: Can Xie

Shimada I "Two-state conformations in the hyarulonan-binding domain regulate CD44 adhesiveness under flow condition"

Shimada I“透明质酸结合域中的两种状态构象在流动条件下调节 CD44 粘附性”

• 发表时间: 2010
• 期刊: Structure 18
• 作者: Ogino S;Nishida N;Umemoto R;Suzuki M;Takeda M;Terasawa H;Kitayama J;Matsumoto M;Hayasaka H;Miyasaka M;Shimada I
• 通讯作者: Shimada I

Observation of NMR Signals from Proteins Introduced into Living Mammalian Cells by Reversible Membrane Permeabilization Using a Pore-Forming Toxin, Streptolysin O

• DOI: 10.1021/ja904407w
• 发表时间: 2009-08-12
• 期刊: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
• 影响因子: 15
• 作者: Ogino, Shinji;Kubo, Satoshi;Shimada, Ichio
• 通讯作者: Shimada, Ichio

Springer TA "Structure of an integrin with anαI domain, complement receptor type 4"

Springer TA“具有 αI 结构域的整合素结构，补体受体 4 型”

• 发表时间: 2010
• 期刊: EMBO J 29
• 作者: Xie C;Zhu J;Chen X;Mi LZ;Nishida N;Springer TA
• 通讯作者: Springer TA

Shimada I "Observation of NMR signals from proteins introduced into living mammalian cells by reversible membrane permeabilization using a pore-forming toxin, Streptolysin O"

Shimada I“使用成孔毒素链球菌溶血素 O 通过可逆膜透化引入活哺乳动物细胞的蛋白质的 NMR 信号观察”

• 发表时间: 2009
• 期刊: J Am Chem Soc 131
• 作者: Ogino S;Kubo S;Umemoto R;Huang S;Nishida N;Shimada I
• 通讯作者: Shimada I