Subunit composition of plasma von Willebrand factor. Cleavage is present in normal individuals, increased in IIA and IIB von Willebrand disease, but minimal in variants with aberrant structure of individual oligomers (types IIC, IID, and IIE).
Subunit composition of plasma von Willebrand factor. Cleavage is present in normal individuals, increased in IIA and IIB von Willebrand disease, but minimal in variants with aberrant structure of individual oligomers (types IIC, IID, and IIE).
复制标题
血浆冯维勒布兰德因子的亚基组成。
DOI：
10.1172/jci112394
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Nannini,LH
中科院分区：
文献类型：
--
作者：
Zimmerman,TS;Dent,JA;Ruggeri,ZM;Nannini,LH
我们评估了血浆von Willebrand因子(VWF)的亚基组成，发现有证据表明，在正常人中存在切割，在IIA和IIB von Willebrand病(VWD)中增加，但在个别寡聚体结构异常的变异体中减少或不存在。在蛋白水解酶抑制剂存在下，用单抗免疫亲和层析法从血浆中快速纯化VWF。经还原和含有十二烷基硫酸钠的5%聚丙烯酰胺凝胶电泳法，在25例正常人血浆vWF中检测到189、176和140kD的片段以及主要的225 kD亚基。用55株抗vWF单抗免疫印迹、125I兔抗鼠抗体和放射自显影检测vWF多肽。在5名IIA型和5名IIB型vWD患者中，176和140-kD片段的比例相对于完整的225-kD亚基的比例增加，这是通过切除每一条带和定量结合的放射性来确定的。相反，这些片段要么是检测不到的，要么是在单个寡聚体结构异常的变异体(IIC和IID型，以及一个新的变异体，IIE型vWD)中仅以微量存在。这里报道的结果提供了证据，在这两组变异体中缺少大的vWF多聚体是由不同的机制造成的。此外，他们还证明了血浆vWF亚单位的部分切割是正常事件。
We have evaluated the subunit composition of plasma von Willebrand factor (vWF) and found evidence that cleavage is present in normal individuals, increased in IIA and IIB von Willebrand disease (vWD), but decreased or absent in variants with aberrant structure of individual oligomers. vWF was rapidly purified from plasma on an analytical scale by monoclonal antibody immunoaffinity chromatography in the presence of protease inhibitors. After reduction and electrophoresis in 5% polyacrylamide gels containing sodium dodecyl sulfate, fragments of 189, 176, and 140 kD, as well as the predominant 225-kD subunit, were identified in plasma vWF from 25 normal individuals. The vWF polypeptides were detected by immunoblotting with a mixture of 55 anti-vWF monoclonal antibodies followed by 125I-rabbit anti-mouse antibody and autoradiography. In five individuals with type IIA and five individuals with type IIB vWD, the proportions of 176 and 140-kD fragments were increased relative to the intact 225-kD subunit, as determined by excising each band and quantitating incorporated radioactivity. In contrast, these fragments were either not detectable or were present in only trace amounts in variants with abnormal structure of individual oligomers (types IIC and IID, and a new variant, type IIE vWD). The results reported here provide evidence that absence of large vWF multimers in these two groups of variants results from different mechanisms. In addition, they demonstrate that partial cleavage of the plasma vWF subunit is a normal event.Images