van der Schoot C E, Wester M, Von Dem Borne A E, Huisman H G
Br J Haematol. 1986 Dec;64(4):715-23. doi: 10.1111/j.1365-2141.1986.tb02233.x.
The glycoprotein localization of the platelet-specific antigens Zwa, Zwb and Baka and their presence on tryptic fragments of glycoproteins was studied by immunoblotting. Human platelets were solubilized and pre-cleared from platelet-associated IgG. The glycoproteins were separated on SDS polyacrylamide gels, transferred to nitrocellulose and incubated with platelet antibodies, followed by 125I-radiolabelled anti-human Ig antibodies. Glycoprotein IIb/IIIa were isolated from platelet lysates by immuno-affinity chromatography. These proteins were subjected to trypsin digestion, and then used for the immunoblot procedure with platelet antibodies. A glycoprotein specifically reacting with either anti-Zwa or anti-Zwb was found, with an apparent molecular weight of 88 kDa. This protein co-migrated, and was probably identical with, glycoprotein IIIa. After trypsin digestion the smallest fragment, reactive with IgG anti-Zwa or IgM anti-Zwb, had a molecular weight of approximately 23 kDa. IgG anti-Baka and anti-Leka antibodies reacted with a protein of 130 kDa from platelets of Bak(a+) donors. This protein was identified as glycoprotein IIb.
通过免疫印迹法研究了血小板特异性抗原Zwa、Zwb和Baka的糖蛋白定位及其在糖蛋白胰蛋白酶片段上的存在情况。将人血小板溶解，并从血小板相关IgG中预先清除。糖蛋白在SDS聚丙烯酰胺凝胶上分离，转移至硝酸纤维素膜，与血小板抗体孵育，然后用125I放射性标记的抗人Ig抗体孵育。通过免疫亲和色谱从血小板裂解物中分离糖蛋白IIb/IIIa。将这些蛋白进行胰蛋白酶消化，然后用于与血小板抗体的免疫印迹程序。发现一种与抗Zwa或抗Zwb特异性反应的糖蛋白，其表观分子量为88 kDa。该蛋白迁移位置相同，可能与糖蛋白IIIa相同。胰蛋白酶消化后，与抗Zwa IgG或抗Zwb IgM反应的最小片段分子量约为23 kDa。抗Baka IgG和抗Leka抗体与Bak(a+)供体血小板中一种130 kDa的蛋白反应。该蛋白被鉴定为糖蛋白IIb。
