Gurd J W, Evans W H
Can J Biochem. 1976 May;54(5):477-80. doi: 10.1139/o76-068.
Following electrophoresis of ovalbumin in sodium dodecyl sulfate (SDS) this glycoprotein bound 125I-labelled concanavalin A (Con A). The reaction was specific and proportional to the amount of glycoprotein present on the gel. This technique was used to study the Con-A-binding glycoproteins of liver cell surfaces. Mouse liver plasma membranes were purified and subfractionated to yield two fractions corresponding to the bile canalicular surface and the surface between adjacent hepatocytes (Evans, W.H. (1970) Biochem. J. 116, 833-842). Both fractions bound 125I-labelled Con A, the former binding two to three times more lectin than the latter. Following SDS gel electrophoresis individual membrane glycoproteins reacted with 125I-labelled Con A. Both membrane subfractions yielded qualitatively similar Con A binding profiles, seven binding proteins being present in each. The results are consistent with a generally uniform distribution of glycoproteins over the hepatocyte surface. The reaction of lectins with glycoproteins following SDS gel electrophoresis should find general application in the study of membrane composition.
在十二烷基硫酸钠（SDS）中对卵清蛋白进行电泳后，这种糖蛋白与125I标记的伴刀豆球蛋白A（Con A）结合。该反应具有特异性，且与凝胶上存在的糖蛋白量成正比。此技术用于研究肝细胞表面的Con A结合糖蛋白。纯化小鼠肝细胞膜并进行亚分级分离，得到对应胆小管表面和相邻肝细胞之间表面的两个级分（埃文斯，W.H.（1970年）《生物化学杂志》116卷，833 - 842页）。两个级分均与125I标记的Con A结合，前者结合的凝集素比后者多两到三倍。SDS凝胶电泳后，单个膜糖蛋白与125I标记的Con A发生反应。两个膜亚级分产生的Con A结合谱在质量上相似，每个级分中都存在七种结合蛋白。结果与糖蛋白在肝细胞表面普遍均匀分布一致。SDS凝胶电泳后凝集素与糖蛋白的反应在膜组成研究中应具有广泛应用。
