The importance of vicinal cysteines, C1669 and C1670, for von Willebrand factor A2 domain function
The importance of vicinal cysteines, C1669 and C1670, for von Willebrand factor A2 domain function
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DOI：
10.1182/blood-2009-12-257949
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发表时间：
2010-06-10
期刊：
影响因子：
20.3
通讯作者：
Crawley, James T. B.
中科院分区：
文献类型：
--
作者：
Luken, Brenda M.;Winn, Luke Y. N.;Crawley, James T. B.
血管性血友病因子（VWF）A2晶体结构揭示了C1669和C1670之间存在罕见的邻位二硫键，预计会影响ADAMTS 13蛋白水解所需的结构域展开。我们制备了具有（A2-VicCC，残基1473-1670）和不具有邻位二硫键（A2-Δ CC，残基1473-1668）的VWF A2结构域片段。与A2-Delta CC相比，A2-VicCC表现出受损的蛋白水解，并且与ADAMTS 13的结合也减少。圆二色性研究表明，A2-VicCC比A2-Delta CC更耐热去折叠。全长VWF中C1669/C1670的突变导致对ADAMTS 13切割的敏感性显著增加，证实了成对的邻近半胱氨酸在VWF A2结构域稳定中的重要作用。（血。2010;115（23）：4910-4913）
The von Willebrand factor (VWF) A2 crystal structure has revealed the presence of a rare vicinal disulfide bond between C1669 and C1670, predicted to influence domain unfolding required for proteolysis by ADAMTS13. We prepared VWF A2 domain fragments with (A2-VicCC, residues 1473-1670) and without the vicinal disulfide bond (A2-Delta CC, residues 1473-1668). Compared with A2-Delta CC, A2-VicCC exhibited impaired proteolysis and also reduced binding to ADAMTS13. Circular dichroism studies revealed that A2-VicCC was more resistant to thermal unfolding than A2-Delta CC. Mutagenesis of C1669/C1670 in full-length VWF resulted in markedly increased susceptibility to cleavage by ADAMTS13, confirming the important role of the paired vicinal cysteines in VWF A2 domain stabilization. (Blood. 2010;115(23):4910-4913)