论文部分内容阅读
目的:研究富含脯氨酸的肌醇多磷酸5-磷酸酶(proline-rich inositol polyphosphate 5-phosphatase,PIPP)在组氨酸557位点突变为丙氨酸(FLAG-PIPPH557A)时对小鼠受精卵中磷酸化苏氨酸蛋白激酶(pAkt)的影响。方法:将FLAG-PIPPH557A、FLAG-PIPP以及FLAG vector重组质粒转染到小鼠受精卵,用蛋白印记、免疫荧光染色以及活性检测方法分析FLAG-PIPPH557A对小鼠受精卵中丝氨酸(Ser)473位点磷酸化的苏氨酸蛋白激酶(pAkt Ser473)表达、定位以及活性的影响。结果:FLAG-PIPPH557A在小鼠受精卵中对pAkt Ser473的表达和活性的调节作用明显低于FLAG-PIPP的作用,两者间有显著差异(P<0.001);而且在小鼠受精卵中FLAG-PIPPH557A不能调节pAktSer473在细胞膜的定位。结论:PIPPH557A在小鼠1-细胞期受精卵中可以通过改变PIPP的活性,影响pAkt Ser473的表达、定位以及活性。
Objective: To investigate the effect of proline-rich inositol polyphosphate 5-phosphatase (PIPP) on the mutation of histidine 557 to alanine (FLAG-PIPPH557A) Effects of phosphorylated threonine protein kinase (pAkt) in fertilized eggs. Methods: FLAG-PIPPH557A, FLAG-PIPP and FLAG vector were transfected into mouse zygotes, and the effect of FLAG-PIPPH557A on the serine (Ser) 473 of mouse fertilized eggs was analyzed by Western blotting, immunofluorescence and activity assay Effect of the expression, localization and activity of the phosphorylated threonine protein kinase (pAkt Ser473). RESULTS: FLAG-PIPPH557A had a significantly lower effect on the expression and activity of pAkt Ser473 in mouse zygotes than FLAG-PIPP (P <0.001) -PIPPH557A does not regulate the localization of pAktSer473 in the cell membrane. Conclusion: PIPPH557A can affect the expression, localization and activity of pAkt Ser473 by changing the activity of PIPP in mouse 1-cell stage zygotes.