Sawyer J T, Sanford J P, Doyle D
Department of Biological Sciences, State University of New York, Buffalo 14260.
J Biol Chem. 1988 Jul 25;263(21):10534-8.
We have generated antibodies against synthetic peptides which represent the carboxyl terminus of either the major, or the two minor, forms of the rat hepatic lectin which recognizes galactose-terminated glycoproteins (asialoglycoproteins). The antibodies were shown to be specific for the form of the lectin containing the immunizing peptide sequence by the following: reaction with purified lectin after sodium dodecyl sulfate-polyacrylamide gel electrophoresis, immunoprecipitation of sodium dodecyl sulfate-denatured lectin, immunoprecipitation of lectin synthesized in vitro. These antibodies, however, precipitated all three rat hepatic lectin forms from nonionic detergent extracts of hepatocytes labeled with 125I via the lactoperoxidase catalyzed technique. A similar result was obtained if antibody was bound to intact cells prior to solubilization with detergent and collection of the immune complexes. We conclude that at least the plasma membrane-associated fraction of the rat hepatic lectin forms exists as a heterotypic complex.
我们已经制备了针对合成肽的抗体,这些合成肽代表大鼠肝凝集素主要形式或两种次要形式的羧基末端,该凝集素可识别半乳糖末端糖蛋白(去唾液酸糖蛋白)。通过以下方式表明这些抗体对含有免疫肽序列的凝集素形式具有特异性:在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳后与纯化的凝集素反应、对十二烷基硫酸钠变性的凝集素进行免疫沉淀、对体外合成的凝集素进行免疫沉淀。然而,这些抗体通过乳过氧化物酶催化技术从用¹²⁵I标记的肝细胞的非离子去污剂提取物中沉淀出所有三种大鼠肝凝集素形式。如果在用去污剂溶解并收集免疫复合物之前将抗体与完整细胞结合,也会得到类似的结果。我们得出结论,至少大鼠肝凝集素形式的质膜相关部分以异型复合物的形式存在。