Covalent regulation of ULVWF string formation and elongation on endothelial cells under flow conditions.
Covalent regulation of ULVWF string formation and elongation on endothelial cells under flow conditions.
复制标题
在流动条件下，对内皮细胞上ULVWF弦线形成和伸长的共价调节。
DOI：
10.1111/j.1538-7836.2008.02991.x
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发表时间：
2008-07
影响因子：
10.4
通讯作者：
Dong, J. -F.
中科院分区：
文献类型：
--
作者：
Li, Y.;Choi, H.;Zhou, Z.;Nolasco, L.;Pownall, H. J.;Voorberg, J.;Moake, J. L.;Dong, J. -F.
粘附配体血管性血友病因子（VWF）是一种多聚体糖蛋白，介导血小板粘附到暴露的内皮下。在内皮细胞上，新鲜释放的超大（UL）VWF多聚体形成血小板粘附的长串样结构。在硫醇封端的N-乙基马来酰亚胺（NEM）存在下，研究了ULVWF串的形成和延伸。通过马来酰亚胺-PEO 2-生物素标记和巯基色谱法测定ULVWF和血浆VWF多聚体中巯基的存在。最后，在无细胞和无酶系统中检查了（UL）VWF的共价再多聚化。我们发现纯化的血浆VWF多聚体（但当硫醇被阻断时效率显著降低）在流动条件下粘附并延长ULVWF串。通过用NEM阻断VWF上的硫醇，ULVWF串的形成和繁殖呈剂量依赖性地减少，表明ULVWF串是通过灌注的VWF与锚定到内皮细胞的ULVWF的共价缔合形成的。通过VWF多聚体中游离巯基的存在和（UL）VWF共价再多聚化的能力，使得缔合成为可能。这些数据提供了血栓形成ULVWF串在内皮细胞上形成和延长的机制。这一机制表明ULVWF的巯基-二硫键状态调节内皮细胞上的弦的粘附特性。
The adhesion ligand von Willebrand factor (VWF) is a multimeric glycoprotein that mediates platelet adhesion to exposed subendothelium. On endothelial cells, freshly released ultra-large (UL)VWF multimers form long string-like structures to which platelets adhere. The formation and elongation of ULVWF strings were studied in the presence of the thiol-blocking N-ethylmaleimide (NEM). The presence of thiols in ULVWF and plasma VWF multimers was determined by maleimide-PEO2-Biotin labeling and thiol-chromatography. Finally, covalent re-multimerization of (UL)VWF was examined in a cell- and enzyme-free system. We found that purified plasma VWF multimers (but significantly less efficient when thiols are blocked) adhere to and elongate ULVWF strings under flow conditions. The formation and propagation of ULVWF strings were dose-dependently reduced by blocking thiols on VWF with NEM, indicating that ULVWF strings are formed by the covalent association of perfused VWF to ULVWF anchored to endothelial cells. The association is made possible by the presence of free thiols in VWF multimers and by the ability of (UL)VWF to covalently re-multimerize. The data provide a mechanism by which the thrombogenic ULVWF strings are formed and elongated on endothelial cells. This mechanism suggests that the thiol-disulfide state of ULVWF regulates the adhesion properties of strings on endothelial cells.
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影响因子：
20.3
作者：
Kenny, D;Jónsson, OG;Montgomery, RR
通讯作者：
Montgomery, RR
DOI：
10.1073/pnas.012459599
发表时间：
2002-01-08
影响因子：
11.1
作者：
Savage, B;Sixma, JJ;Ruggeri, ZM
通讯作者：
Ruggeri, ZM
影响因子：
4.8
作者：
Burgess, JK;Hotchkiss, KA;Hogg, PJ
通讯作者：
Hogg, PJ
影响因子：
20.3
作者：
Arya, M;Anvari, B;López, JA
通讯作者：
López, JA
影响因子：
3.6
作者：
Jensen, LH;Renodon-Corniere, A;Jensen, PB
通讯作者：
Jensen, PB