A single high-affinity binding site for von Willebrand factor in collagen III, identified using synthetic triple-helical peptides
A single high-affinity binding site for von Willebrand factor in collagen III, identified using synthetic triple-helical peptides
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DOI：
10.1182/blood-2006-03-011965
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发表时间：
2006-12-01
期刊：
影响因子：
20.3
通讯作者：
Farndale, Richard W.
中科院分区：
文献类型：
--
作者：
Lisman, Ton;Raynal, Nicolas;Farndale, Richard W.
血小板粘附至受损血管壁的基本事件是血浆血管性血友病因子（VWF）与内皮下胶原的结合，VWF是一种与血小板糖蛋白Ib α（GPIb α）短暂相互作用的蛋白质，减缓循环血小板以促进通过胶原受体（包括整合素α 2 β 1和GpVI）的牢固粘附。为了定位在胶原结合VWF的网站，我们合成了57个重叠的三螺旋肽，包括整个三螺旋结构域的胶原III。肽23单独结合VWF，具有与天然胶原III相似的亲和力。固定化肽23号在静态和流动条件下支持血小板粘附，该过程被阻止胶原蛋白结合VWF A3结构域的抗体阻断。来自23号的截短和丙氨酸取代的肽与VWF和血小板两者强烈相互作用或缺乏VWF和血小板结合。因此，我们确定了序列RGQOGVMGF（O是羟脯氨酸）作为胶原蛋白III中最小的VWF结合序列。
The essential event in platelet adhesion to the injured blood vessel wall is the binding to subendothelial collagen of plasma von Willebrand factor (VWF), a protein that interacts transiently with platelet glycoprotein Ib alpha (GPIb alpha), slowing circulating platelets to facilitate firm adhesion through collagen receptors, including integrin alpha 2 beta 1 and GpVI. To locate the site in collagen that binds VWF, we synthesized 57 overlapping triple-helical peptides comprising the whole triple-helical domain of collagen III. Peptide no. 23 alone bound VWF, with similar affinity to that of native collagen III. Immobilized peptide no. 23 supported platelet adhesion under static and flow conditions, processes blocked by an antibody that prevents collagen from binding the VWF A3 domain. Truncated and alanine-substituted peptides derived from no. 23 either strongly interacted with both VWF and platelets or lacked both VWF and platelet binding. Thus, we identified the sequence RGQOGVMGF (O is hydroxyproline) as the minimal VWF-binding sequence in collagen III.