Aoyagi T, Adachi K, Halprin K M, Levine V
J Invest Dermatol. 1980 Jun;74(6):421-4. doi: 10.1111/1523-1747.ep12544582.
Incorporation of 32P-orthophosphoric acid into pig epidermal proteins was analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The incorporation was observed in 2 major and 2 minor peaks with approximate molecular weights of 52,000 and 27,000, 17,000, and 12,000 respectively. Treatment with epidermal growth factor (EGF) consistently increased 32P incorporation into all 4 peaks. The stimulation was both time- and dose-dependent, and was not inhibited by the simultaneous addition of cycloheximide. In our previous study (submitted to J Invest Dermatol), we observed that EGF increased the cyclic GMP level in the same pig skin (epidermal) system. The possibility that the increased cyclic GMP leads to the protein phosphorylation observed in the present study was considered but ruled out because (1) the addition of cyclic GMP does not mimic the EGF treatment, (2) simultaneous addition of EGF and cyclic GMP stimulate phosphorylation to the same degree as the addition of EGF alone and (3) EGF stimulates phosphorylation in 20 min whereas the EGF induced increase in the cyclic GMP level takes 60 min.
采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析了32P-正磷酸掺入猪表皮蛋白的情况。观察到掺入出现2个主要峰和2个次要峰，其近似分子量分别为52,000、27,000、17,000和12,000。用表皮生长因子（EGF）处理始终会增加32P掺入所有4个峰中的量。这种刺激具有时间和剂量依赖性，并且不会被同时添加放线菌酮所抑制。在我们之前的研究（已提交给《皮肤病学研究杂志》）中，我们观察到EGF会增加同一猪皮肤（表皮）系统中的环磷酸鸟苷水平。曾考虑过环磷酸鸟苷增加导致本研究中观察到的蛋白质磷酸化的可能性，但由于以下原因而排除：（1）添加环磷酸鸟苷并不能模拟EGF处理；（2）同时添加EGF和环磷酸鸟苷刺激磷酸化的程度与单独添加EGF相同；（3）EGF在20分钟内刺激磷酸化，而EGF诱导的环磷酸鸟苷水平升高需要60分钟。
