Nakaya K, Ushiwata A, Nakajo S, Nakamura Y
J Biochem. 1979 Jan;85(1):183-9. doi: 10.1093/oxfordjournals.jbchem.a132308.
The cell surface proteins of AH-66 hepatoma ascites cells were examined both by fluorescent labeling with cycloheptaamylose-fluorescamine complex and by radioiodination with lactoperoxidase. The labeled proteins were analyzed by measuring the distribution of fluorescence or radioactivity in polyacrylamide gels after electrophoresis. Although the degree of fluorescent labeling with cycloheptaamylose-fluorescamine complex was substantially different from that obtained by lactoperoxidase-catalyzed iodination, proteins with molecular weights of 130,000, 90,000, 86,000, 50,000--60,000, and 37,000 were strongly labeled in common by both reagents. It is suggested that at least these proteins are exposed on the surface of the plasma membranes of AH-66 cells. One of these proteins labeled strongly by both reagents is the major glycoprotein, with an apparent molecular weight of 130,000 in 10% polyacrylamide gels.
采用环庚直链淀粉-荧光胺复合物进行荧光标记以及用乳过氧化物酶进行放射性碘标记,对AH-66肝癌腹水细胞的细胞表面蛋白进行了检测。标记后的蛋白通过测量电泳后聚丙烯酰胺凝胶中荧光或放射性的分布来进行分析。尽管环庚直链淀粉-荧光胺复合物的荧光标记程度与乳过氧化物酶催化碘化法得到的结果有很大差异,但分子量为130,000、90,000、86,000、50,000 - 60,000和37,000的蛋白在两种试剂标记下均被强烈标记。提示至少这些蛋白暴露于AH-66细胞的质膜表面。两种试剂均强烈标记的其中一种蛋白是主要糖蛋白,在10%聚丙烯酰胺凝胶中表观分子量为130,000。