Binding and covalent cross-linking of purified von Willebrand factor to native monomeric collagen.
Binding and covalent cross-linking of purified von Willebrand factor to native monomeric collagen.
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纯化的冯维勒布兰德因子与天然单体胶原蛋白的结合和共价交联。
DOI：
10.1172/jci112608
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Handin,RI
中科院分区：
文献类型：
--
作者：
Bockenstedt,P;McDonagh,J;Handin,RI
我们通过将源自小牛皮肤的酸溶性 I 型和 III 型胶原蛋白吸附到聚苯乙烯微量滴定孔中，并用纯化的人 125I-vWF 孵育孔，分析了粘附糖蛋白、冯维勒布兰德因子 (vWF) 与天然单体胶原单层的相互作用。 125I-vWF 的结合是可饱和的、可逆的、特异性的，并且通过胶原单体的热变性而消除。 5 微克/ml 时结合为半最大，并且在饱和状态下，每微克固定胶原蛋白结合 7.5 ng 125 I-vWF。 125I-vWF 不与涂有其他细胞外基质或血浆蛋白（例如纤连蛋白、纤维蛋白原、明胶或补体第一组分的 q 亚基 (C1q)）的孔结合。此外，结合的 125I-vWF 不能通过添加纤连蛋白或纤维蛋白原从胶原蛋白中置换出来。与因子 XIIIa 孵育后，血浆转谷氨酰胺酶、与胶原蛋白结合的 125I-vWF 不再被 vWF 取代，这表明 vWF 与胶原蛋白单体发生共价交联。在十二烷基硫酸钠存在下，聚丙烯酰胺凝胶电泳也证明了 vWF 与胶原蛋白（但不与纤连蛋白或层粘连蛋白）的 XIIIa 因子依赖性共价交联。
We have analyzed the interaction of the adhesive glycoprotein, von Willebrand factor (vWF), with native monomeric collagen monolayers by adsorbing acid soluble Types I and III collagen derived from calf skin to polystyrene microtiter wells and incubating the wells with purified human 125I-vWF. The binding of 125I-vWF was saturable, reversible, specific, and was abolished by heat denaturation of the collagen monomers. Binding was half-maximal at 5 micrograms/ml, and, at saturation, 7.5 ng 125I-vWF were bound to each microgram of immobilized collagen. 125I-vWF did not bind to wells coated with other extracellular matrix or plasma proteins such as fibronectin, fibrinogen, gelatin, or the q subunit of the first component of complement (C1q). In addition, bound 125I-vWF could not be displaced from collagen by the addition of either fibronectin or fibrinogen. After incubation with Factor XIIIa, plasma transglutaminase, 125I-vWF bound to collagen could no longer be displaced by vWF, which suggests covalent cross-linking of vWF to collagen monomers. Factor XIIIa-dependent covalent cross-linking of vWF to collagen, but not to fibronectin or laminin, was also demonstrated by polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate.Images