甲基硝基亚硝胍处理的vero细胞中磷酸化蛋白的差异显示

Differential display of phosphorylated protein in MNNG treated vero cells

目的：初步了解哺乳类细胞非定标性突变形成是否与以蛋白质磷酸化级联反应为主要形式的细胞信号传导通路有关；方法：采用［３２Ｐ］体内预标记及凝胶双向电泳方法差异显示甲基硝基亚硝胍（ＭＮＮＧ）处理组和对照组磷酸化蛋白，蛋白质免疫印迹杂交试验初步鉴定差异显示蛋白斑点性质；结果：在处理组发现两个与对照组不同的蛋白斑点，分子量分别在６８×１０３和４３×１０３附近，与抗酪氨酸磷酸化蛋白抗体未发生阳性反应；结论：根据差异显示蛋白斑点的分子量和印迹杂交试验结果，初步认为所见蛋白斑点可能为丝／苏氨酸磷酸化蛋白。

AIM:To explore the potential relationship between nontargeted mutagenesis and the cascades of protein phosphorylation of signal transduction pathways in cells. METHOD:Differential display of phosphorylated protein in cells treated with MNNG (methylnitronitrosoguanidine) and those with dimethyl sulfoxide (control) were performed with [ 32 P] incorporation in vivo ; Western blotting test was employed to identify the property of the protein. RESULTS:There are two differential displayed phosphorylated proteins reacted negatively with antiphosphotyrosine in MNNG treatment cells. CONCLUSION:The molecular weight of two proteins and the results of Western blot suggested that the two differential displayed phosphorylated proteins may be Ser/Thr phosphoproteins and probably members of the MAPK.