Decker S J
Mol Cell Biol. 1984 Apr;4(4):571-5. doi: 10.1128/mcb.4.4.571-575.1984.
The biosynthesis and posttranslational metabolism of the epidermal growth factor (EGF) receptor were examined in the A431 human epidermoid carcinoma cell line. Polyclonal antibody against the receptor specifically immunoprecipitated two [35S]methionine-labeled proteins of Mr = 160,000 and 170,000. Pulse chase experiments showed the Mr = 160,000 protein to be a precursor of the Mr = 170,000 protein. Preincubation with tunicamycin resulted in immunoprecipitation of a single band of Mr = 130,000, whereas monensin inhibited maturation to the Mr = 170,000 form. Digestion of the Mr = 160,000 and 170,000 proteins with endoglycosidase H resulted in the appearance of Mr = 130,000 and 165,000 proteins, respectively. Prolonged pulse-chase experiments indicated that the half-life of the receptor is ca. 20 h in the absence of EGF and 5 h in the presence of EGF. Approximately three- to five-fold more phosphate is incorporated into the mature receptor upon addition of EGF, due primarily to increases in levels of phosphotyrosine and phosphoserine. Phosphate was also present on the Mr = 160,000 protein and the Mr = 130,000 protein found in the presence of tunicamycin.
在A431人表皮样癌细胞系中研究了表皮生长因子(EGF)受体的生物合成和翻译后代谢。针对该受体的多克隆抗体特异性免疫沉淀出两种分子量分别为160,000和170,000的[35S]甲硫氨酸标记蛋白。脉冲追踪实验表明,分子量为160,000的蛋白是分子量为170,000蛋白的前体。用衣霉素预孵育导致免疫沉淀出一条分子量为130,000的条带,而莫能菌素抑制了向分子量为170,000形式的成熟过程。用内切糖苷酶H消化分子量为160,000和170,000的蛋白分别产生了分子量为130,000和165,000的蛋白。延长的脉冲追踪实验表明,在没有EGF的情况下,受体的半衰期约为20小时,在有EGF的情况下为5小时。添加EGF后,成熟受体中掺入的磷酸盐增加约三到五倍,这主要是由于磷酸酪氨酸和磷酸丝氨酸水平的增加。在衣霉素存在下发现的分子量为160,000的蛋白和分子量为130,000的蛋白上也存在磷酸盐。