ANALYSIS OF THE GLYCOPROTEIN AND PROTEIN-COMPOSITION OF BERNARD-SOULIER PLATELETS BY SINGLE AND TWO-DIMENSIONAL SODIUM DODECYL SULFATE-POLYACRYLAMIDE GEL-ELECTROPHORESIS
ANALYSIS OF THE GLYCOPROTEIN AND PROTEIN-COMPOSITION OF BERNARD-SOULIER PLATELETS BY SINGLE AND TWO-DIMENSIONAL SODIUM DODECYL SULFATE-POLYACRYLAMIDE GEL-ELECTROPHORESIS
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DOI：
10.1172/jci110172
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发表时间：
1981-01-01
影响因子：
15.9
通讯作者：
CAEN, JP
中科院分区：
文献类型：
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作者：
NURDEN, AT;DUPUIS, D;CAEN, JP
以前的报告描述了关于Bernard-Soulier血小板的糖蛋白(GP)和蛋白质组成的相互矛盾的结果。针对这一争议，采用改良的十二烷基硫酸钠(SDS)-聚丙烯酰胺凝胶电泳法对4例Bernard-Soulier患者的血小板进行了分析。每个患者的血小板都没有膜糖蛋白Ib的糖类染色。清晰地检测到膜GP IIb、IIIa和IIIb的主要高碘酸盐-希夫染色条带，并通过双向SDS-聚丙烯酰胺凝胶电泳证实了它们的存在。Bernard-Soulier血小板的蛋白质含量增加了2-4倍。用7-12%的丙烯酰胺梯度凝胶分析它们的蛋白质组成，显示出正常的多肽图谱。采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳法和放射自显影技术对Bernard-Soulier血小板膜表面蛋白进行125I标记。没有检测到Ib位置的标记，而其他主要的膜GP，包括Ia和IIa，正常定位。与之相反，用高碘酸盐-席夫染色和放射自显影技术分析在乳过氧化物酶催化的碘化前用神经氨酸酶处理过的正常人血小板，可以清楚地检测到GP Ib。从患者红细胞膜分离的糖蛋白图谱中未发现异常。只有严重的分子异常或可能的GP Ib缺失才能解释Bernard-Soulier综合征中这一主要的血小板损害。
Previous reports have described conflicting results concerning the glycoprotein (GP) and protein composition of Bernard-Soulier platelets. In view of this controversy an analysis was made of the platelets of 4 Bernard-Soulier patients using improved single and 2-dimensional sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis procedures. An absence of staining for carbohydrate of membrane GP Ib was characteristic for the platelets of each patient. Major periodate-Schiff staining bands corresponding to membrane GP IIb, IIIa and IIIb were clearly detected and their presence was confirmed by 2-dimensional SDS-polyacrylamide gel electrophoresis. The protein content of the Bernard-Soulier platelets was increased 2- to 4-fold. Analysis of their protein composition using 7-12% acrylamide gradient gels showed normal polypeptide profiles. Lactoperoxidase-catalyzed 125I-labeling of the Bernard-Soulier platelet surface proteins was followed by SDS-polyacrylamide gel electrophoresis and autoradiography. No labeling in the Ib position was detected whereas the other major membrane GP, including Ia and IIa, were normally located. In contrast, GP Ib was clearly detected by periodate-Schiff staining and autoradiography when normal human platelets that were exhaustively treated with neuraminidase before the lactoperoxidase-catalyzed iodination were analyzed. No abnormalities were detected in the GP patterns of membranes isolated from the patients'' erythrocytes. Only a severe molecular abnormality or possible deletion of GP Ib could account for this major platelet lesion in the Bernard-Soulier syndrome.