Smith M R, Liu Y L, Kim H, Rhee S G, Kung H F
Biological Carcinogenesis and Development Program, National Cancer Institute-Frederick Cancer Research Facility, MD 21701.
Science. 1990 Mar 2;247(4946):1074-7. doi: 10.1126/science.2408147.
Several immunologically distinct isozymes of inositol phospholipid-specific phospholipase C (PLC) have been purified from bovine brain. Murine NIH 3T3 fibroblasts were found to express PLC-gamma, but the expression of PLC-beta was barely detectable by radioimmunoassay or protein immunoblot. A mixture of monoclonal antibodies was identified that neutralizes the biological activity of both endogenous and injected purified PLC-gamma. When co-injected with oncogenic Ras protein or PLC-gamma, this mixture of antibodies inhibited the induction of DNA synthesis that characteristically results from the injection of these proteins into quiescent 3T3 cells. However, when oncogenic Ras protein or PLC-gamma was co-injected with a neutralizing monoclonal antibody to Ras, only the DNA synthesis induced by the Ras protein was inhibited--that induced by PLC was unaffected. These results suggest that the Ras protein is an upstream effector of PLC activity in phosphoinositide-specific signal transduction and that PLC-gamma activity is necessary for Ras-mediated induction of DNA synthesis.
已经从牛脑中纯化出了几种免疫特性不同的肌醇磷脂特异性磷脂酶C(PLC)同工酶。发现小鼠NIH 3T3成纤维细胞表达PLC-γ,但通过放射免疫测定或蛋白质免疫印迹几乎检测不到PLC-β的表达。鉴定出一种单克隆抗体混合物,它能中和内源性和注射的纯化PLC-γ的生物活性。当与致癌性Ras蛋白或PLC-γ共同注射时,这种抗体混合物抑制了将这些蛋白注射到静止的3T3细胞中所特有的DNA合成诱导。然而,当致癌性Ras蛋白或PLC-γ与针对Ras的中和单克隆抗体共同注射时,仅由Ras蛋白诱导的DNA合成受到抑制,而由PLC诱导的DNA合成不受影响。这些结果表明,在磷酸肌醇特异性信号转导中,Ras蛋白是PLC活性的上游效应物,并且PLC-γ活性是Ras介导的DNA合成诱导所必需的。