Thiery J P, Brackenbury R, Rutishauser U, Edelman G M
J Biol Chem. 1977 Oct 10;252(19):6841-5.
The aggregation of cells from dissociated neural retinas of chick embryos can be inhibited by antibodies prepared against whole retinal cells. In order to identify the antigens involved, substances released by retinal tissues in culture were tested for their ability to neutralize specifically the inhibition by antibody of cell adhesion. Using this assay, three active polypeptides from the culture supernatant were purified 500-fold by gel filtration and polyacrylamide gel electrophoresis. Rabbit antibodies prepared against these purified supernatant activities inhibited cell adhesion and reacted only with the three polypeptides. Immunoprecipitation by the specific antibodies of 3H-labeled proteins from a detergent extract of embryonic retinal cell membranes yielded a polypeptide having a Mr of 140,000 in sodium dodecyl sulfate. This precipitation was inhibited in the presence of the three culture supernatant polypeptides that had activity, suggesting that they contained antigenic determinants in common with the 140,000 Mr surface component. They therefore represent all or parts of this cell surface molecule that were released into solution during tissue culture. The data are consistent with the hypothesis that the 140,000 Mr polypeptide is intimately involved in initial adhesion among neural cells.
针对鸡胚神经视网膜解离细胞的聚集,可被针对全视网膜细胞制备的抗体所抑制。为了鉴定其中涉及的抗原,对培养的视网膜组织释放的物质进行了测试,以检测它们特异性中和抗体对细胞黏附抑制作用的能力。利用该检测方法,通过凝胶过滤和聚丙烯酰胺凝胶电泳,从培养上清液中纯化出三种活性多肽,纯化倍数达500倍。针对这些纯化的上清液活性物质制备的兔抗体抑制细胞黏附,且仅与这三种多肽发生反应。用特异性抗体对胚胎视网膜细胞膜去污剂提取物中的3H标记蛋白进行免疫沉淀,在十二烷基硫酸钠中产生一种分子量为140,000的多肽。在具有活性的三种培养上清液多肽存在的情况下,这种沉淀受到抑制,这表明它们含有与140,000分子量表面成分共同的抗原决定簇。因此,它们代表了在组织培养过程中释放到溶液中的该细胞表面分子的全部或部分。这些数据与以下假设一致,即140,000分子量的多肽与神经细胞之间的初始黏附密切相关。