Aoyagi T
Hokkaido Igaku Zasshi. 1981 May;56(3):319-33.
Incubation of pig skin slices with epidermal growth factor (EGF) caused an increase in intracellular cyclic GMP concentration of pig epidermis. A significant increase was found after 1 hr of incubation and reached a peak by 6 hrs. EGF caused no change in the cyclic AMP level, nor did it affect epinephrine-induced cyclic AMP responses. EGF also stimulated protein phosphorylation in pig epidermis. The stimulation was both time and dose dependent, and was not inhibited by the simultaneous addition of cycloheximide. The addition of cyclic GMP did not mimic the EGF treatment and EGF stimulated protein phosphorylation in 20 minutes. Phosphoproteins stimulated by EGF were identified by SDS-PAGE and autoradiographic analysis. The identified phosphoproteins were fibrous keratin proteins (MW. 65,000, 60,000, 56,000 and 51,000) and soluble protein (MW. 30,000). Calcium ion potentiated the phosphorylation of keratin proteins by EGF. Ionophore A23187 plus calcium ion also enhanced the phosphorylation of keratin proteins.
用表皮生长因子(EGF)孵育猪皮切片会导致猪表皮细胞内环状鸟苷酸(cGMP)浓度升高。孵育1小时后发现有显著升高,并在6小时时达到峰值。EGF不会改变环磷酸腺苷(cAMP)水平,也不影响肾上腺素诱导的cAMP反应。EGF还能刺激猪表皮中的蛋白质磷酸化。这种刺激具有时间和剂量依赖性,并且不会被同时添加的环己酰亚胺抑制。添加cGMP并不能模拟EGF处理,且EGF在20分钟内就能刺激蛋白质磷酸化。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和放射自显影分析鉴定了受EGF刺激的磷蛋白。鉴定出的磷蛋白为纤维角蛋白(分子量分别为65,000、60,000、56,000和51,000)和可溶性蛋白(分子量为30,000)。钙离子增强了EGF对角蛋白的磷酸化作用。离子载体A23187加钙离子也增强了角蛋白的磷酸化作用。