Kung H F, Smith M R, Bekesi E, Manne V, Stacey D W
Exp Cell Res. 1986 Feb;162(2):363-71. doi: 10.1016/0014-4827(86)90341-1.
The transforming activities of p21 ras proteins have been determined by micro-injection of these proteins into NIH3T3 cells. In order to facilitate functional studies on the effect of ras proteins on malignant transformation and normal cellular growth, analysis has been made with three monoclonal antibodies (YA6-172, Y13-238 and Y13-259) as originally reported by Furth et al. (J virol 43 (1982) 294). Purified immunoglobulin of Y13-259 has the highest titer of binding to bacterially synthesized p21 ras proteins. Experimental analyses indicate that only Y13-259 antibody will neutralize the transforming activity of the co-injected bacterially synthesized ras protein and the neutralization effect was blocked by co-injection of excess ras protein. In addition, micro-injection of Y13-259 immunoglobulin into transformed NIH3T3 cells (obtained by DNA transfection of NIH3T3 cells with molecularly cloned ras gene) reversed their transformed phenotypes. These results indicate that both bacterially synthesized p21 ras proteins and the natural ras proteins produced in NIH3T3 cells were neutralized by Y13-259 antibody.
通过将p21 ras蛋白显微注射到NIH3T3细胞中，已确定了这些蛋白的转化活性。为便于对ras蛋白对恶性转化和正常细胞生长的影响进行功能研究，已如Furth等人最初报道的那样（《病毒学杂志》43(1982)294），使用三种单克隆抗体（YA6-172、Y13-238和Y13-259）进行了分析。Y13-259的纯化免疫球蛋白与细菌合成的p21 ras蛋白的结合效价最高。实验分析表明，只有Y13-259抗体能中和共注射的细菌合成ras蛋白的转化活性，且过量ras蛋白的共注射会阻断这种中和作用。此外，将Y13-259免疫球蛋白显微注射到转化的NIH3T3细胞（通过用分子克隆的ras基因对NIH3T3细胞进行DNA转染获得）中，可逆转其转化表型。这些结果表明，Y13-259抗体既能中和细菌合成的p21 ras蛋白，也能中和NIH3T3细胞中产生的天然ras蛋白。
