Isolation and immunologic characterization of the human platelet alloantigen, P1A1.
Isolation and immunologic characterization of the human platelet alloantigen, P1A1.
复制标题
人血小板同种抗原 P1A1 的分离和免疫学特征。
DOI：
10.1016/0161-5890(79)90100-7
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发表时间：
1979
影响因子：
3.6
通讯作者：
R. Aster
中科院分区：
文献类型：
--
作者：
T. Kunicki;R. Aster
格兰兹曼血栓减少症（血小板功能的先天性障碍）患者的血小板缺乏膜同种异体抗原。P1a1 (Zwa)。存在于几乎所有正常人的血小板中。因为血栓性血小板也缺乏两种膜糖蛋白，称为IIb和IIIa。有人认为这些糖蛋白中的一种或两种可能携带p1a1抗原决定因子。为了解决这一问题，我们采用血小板膜脱氧胆酸钠提取物的序列凝集素亲和层析法，乳酸过氧化物酶碘化完整血小板的Nonidet P40提取物的间接免疫沉淀法，以及制备十二烷基硫酸钠-聚丙烯酰胺凝胶电泳的增溶膜制剂分离p1a1抗原。通过三种分离方法中的每一种，p1a1抗原标记都被证明与糖蛋白IIIa相关。对蛋白水解酶、二硫反应剂和温度对p1a1活性的影响以及各种糖对p1a1 -抗p1a1反应的影响的研究表明，p1a1抗原位于胰蛋白酶易裂解的分子部分，而不是胰凝乳蛋白酶。菠萝蛋白酶或木瓜蛋白酶需要完整的二硫键才能完全表达，这可能是由GPIIIa中的多肽序列决定的。几乎所有血小板的p1a1活性都位于外质膜上。p1a1似乎是第一个被分配到特定血小板膜成分的同种异体抗原，也是第三个被分配到人类细胞膜蛋白的由肽决定的同种异体抗原。
Platelets from patients with Glanzmann's thrombasthenia, acongential disorder of platelet function, are deficient in the membrane alloantigen. P1a1(Zwa). present on platelets of nearly all normal subjects. Because thrombasthenic platelets are also deficient in two membrane glycoproteins, designated IIb and IIIa. it has been suggested that one or both of these glycoproteins may carry the P1a1antigenic determinant. To address this question, the P1a1antigen was isolated by sequential lectin affinity chromatography of sodium deoxycholate extracts of platelet membranes, by indirect immunoprecipitation of Nonidet P40 extracts of lactoperoxidase-iodinated intact platelets, and by preparative sodium dodecyl sulfate-polyacrylamide gel electrophoresis of solubilized membrane preparations. By each of the three separatory procedures, the P1a1antigenic marker was shown to be associated with glycoprotein IIIa.Studies of the effects of proteolytic enzymes, disulfide-reactive agents and temperature on P1a1activity and of the effect of various sugars on the P1a1-anti-P1a1reaction indicate that the P1a1antigen, situated on a portion of the molecule susceptible to cleavagein situby trypsin, but not by chymotrypsin. bromelain, or papain, requires intact disulfide bonds for its full expression, and is probably determined by a polypeptide sequence in GPIIIa. Virtually all of the P1a1activity of platelets appears to be located on the external plasma membrane.P1a1appears to be the first alloantigen to be assigned to a specific platelet membrane constituent and the third peptide-determined alloantigen to be assigned to a human cell membrane protein.