Skoog V T, Nilsson S F, Weber T H
Scand J Immunol. 1980;11(4):369-76. doi: 10.1111/j.1365-3083.1980.tb00002.x.
To compare the receptor patterns for mitogenic and non-mitogenic substances, surface glycoproteins of human lymphocytes were labelled with the lactoperioxidase-catalysed iodination technique and with a galactose oxidase-tritiated sodium borohydride technique. Labelled cells were detergent-solubilized, and the lysates were allowed to react with insolubilized purified mitogenic lectins, phytohaemagglutinin, leucoagglutinin and an insolubilized non-mitogenic lectin, oxidized leucoagglutinin. Lectin-reactive proteins were eluted with sodium dodecyl sulphate (SDS) buffer. Cell membrane components reactive with anti-lymphocyte globulin (ALG) were retrieved by indirect immunoprecipitation with protein-A-bearing staphylococcus Cowan I strain (SaCI). Lectin- and ALG-reactive proteins were analysed by SDS polyacrylamide gel electrophoresis. Iodinated glycoproteins regularly showed four major components with molecular weights of 120,000, 70,000, 60,000 and 43,000 daltons, respectively, on 7% gels. An additional broad peak in the molecular weight range 20,000--35,000 daltons was found on 10% gels. Tritiated glycoproteins also showed four major components with MW 120,000, 70,000, 60,000 AND 42,000, RESPECTIVely, which reacted with lectin and ALG. In addition, ALG reacted with some glycoproteins with MW between 150,000 and 230,000 daltons. On 10% gels additional lectin- and ALG-binding glycoproteins with MW around 30,000 daltons were found. The similarity in structures bound by mitogenic and non-mitogenic substances indicates that lymphocyte activation may depend on some property conferred by the mitogen.
为比较促有丝分裂物质和非促有丝分裂物质的受体模式，采用乳过氧化物酶催化碘化技术和半乳糖氧化酶-氚化硼氢化钠技术对人淋巴细胞的表面糖蛋白进行标记。将标记细胞用去污剂溶解，使裂解物与不溶性纯化促有丝分裂凝集素、植物血凝素、白细胞凝集素以及不溶性非促有丝分裂凝集素氧化白细胞凝集素反应。用十二烷基硫酸钠（SDS）缓冲液洗脱与凝集素反应的蛋白质。通过用含蛋白A的金黄色葡萄球菌考恩I株（SaCI）进行间接免疫沉淀来回收与抗淋巴细胞球蛋白（ALG）反应的细胞膜成分。通过SDS聚丙烯酰胺凝胶电泳分析与凝集素和ALG反应的蛋白质。在7%的凝胶上，碘化糖蛋白通常显示出四个主要成分，分子量分别为120,000、70,000、60,000和43,000道尔顿。在10%的凝胶上发现分子量范围在20,000 - 35,000道尔顿之间有一个额外的宽峰。氚化糖蛋白也显示出四个主要成分，分子量分别为120,000、70,000、60,000和42,000，它们与凝集素和ALG反应。此外，ALG与一些分子量在150,000至230,000道尔顿之间的糖蛋白反应。在10%的凝胶上发现了分子量约为30,000道尔顿的额外的与凝集素和ALG结合的糖蛋白。促有丝分裂物质和非促有丝分裂物质所结合结构的相似性表明淋巴细胞活化可能取决于促有丝分裂原赋予的某些特性。
