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FLAG-PIPP^(H557A)在小鼠1-细胞期受精卵中对PKB/Akt表达的调节作用 被引量：2

Regulative Role of FLAG-PIPP^(H557A) on Phosphorylated Akt at Ser473 in Mouse Fertilized Eggs
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摘要 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 study the regulative role of PIPPH557A（proline-rich inositol polyphosphate 5-phosphatase histidine 557 mutating to alanine） on protein kinase B（PKB/Akt） of mouse fertilized eggs.Methods： With respective transfection of FLAG-PIPPH557A,FLAG-PIPP or FLAG vector to mouse fertilized eggs to detect the expression,location and activity of pAkt Ser473 by Western blotting,indirect immunofluorescence analysis and activity analysis.Results： Effects of catalytically incactive FLAG-PIPPH557A,generated by mutating histidine 557 within the PIPP sequence to alanine,on expression and activity of pAkt Ser473 were lower than those of FLAG-PIPP in mouse fertilized eggs.Their differences were clearly significant（P0.001）.And FLAG-PIPP did not regulate the localization of pAkt Ser473 in cytomembrane.Conclusion： PIPPH557A in 1-cell fertilized eggs may affect the expression,localization and activity of pAktSer473 by changing PIPP activity.
作者 邓欣 潘学风 李亘松 于秉治
机构地区 中国医科大学基础医学院机能实验中心 沈阳医学院沈州医院北院区口腔科 中国医科大学基础医学院生理学教研室 中国医科大学基础医学院生物化学与分子生物学教研室
出处 《生殖与避孕》 CAS CSCD 北大核心 2011年第6期359-363,408,共6页 Reproduction and Contraception
基金 国家自然科学基金项目 项目编号:81070489
关键词 组氨酸557位点突变为丙氨酸的富含脯氨酸的肌醇多磷酸5-磷酸酶(PIPPH557A) 小鼠受精卵 丝氨酸(Ser)473位点磷酸化的苏氨酸蛋白激酶(pAktSer473) proline-rich inositol polyphosphate 5-phosphatase histidine mutating to alanine（PIPPH557A） mouse fertilized eggs pAkt-Ser473
分类号 R339.22 [医药卫生—人体生理学]
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参考文献12

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二级参考文献23

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共引文献8

同被引文献22

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生殖与避孕
2011年 第6期

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