CHARACTERIZATION OF THE UNIQUE MECHANISM MEDIATING THE SHEAR-DEPENDENT BINDING OF SOLUBLE VON-WILLEBRAND-FACTOR TO PLATELETS
CHARACTERIZATION OF THE UNIQUE MECHANISM MEDIATING THE SHEAR-DEPENDENT BINDING OF SOLUBLE VON-WILLEBRAND-FACTOR TO PLATELETS
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DOI：
10.1074/jbc.270.40.23352
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发表时间：
1995-10-06
影响因子：
4.8
通讯作者：
RUGGERI, ZM
中科院分区：
文献类型：
--
作者：
GOTO, S;SALOMON, DR;RUGGERI, ZM
本文研究了异硫氰酸荧光素（FITC）标记的可溶性血管性血友病因子（vWF）与血小板在锥板粘度计中60 °剪切作用下的相互作用机制。使用用荧光珠标准品校准的流式细胞仪计算与悬浮液中的每个血小板相关的分子数。为了验证所使用的方法和试剂，在存在利托那肽或α-凝血酶的情况下评估相同标记的vWF的结合，发现其是可饱和的，在> 90%的血小板上具有窄且对称的分布。正如预期的那样，基本上所有结合的配体在利托那肽存在下仅与血小板膜糖蛋白（GP）Ib α相互作用，在α-凝血酶刺激后与GP IIb-IIIa相互作用。相比之下，只有一小部分（
We have studied the mechanism of interaction between soluble von Willebrand factor (vWF), labeled with fluorescein isothiocyanate (FITC), and platelets exposed 60 shear in a cone-and-plate viscometer. A flow cytometer calibrated with fluorescent bead standards was used 60 calculate the number of molecules associated with each platelet in suspension. To validate the methods and reagents used, binding of the same labeled vWF was assessed in the presence of ristocetin or alpha-thrombin and found to be saturable, with a narrow and symmetric distribution on >90% of the platelets. As expected, essentially all bound ligand interacted exclusively with platelet membrane glycoprotein (GP) Ib alpha in the presence of ristocetin and with GP IIb-IIIa after stimulation with alpha-thrombin. In contrast, only a minor proportion (