PKB/Akt通过Girdin蛋白调控小鼠受精卵微丝聚集  

PKB/Akt regulates the aggregation of actin by Girdin in mouse fertilized eggs

作　　者：武迪迪[1] 张盼盼[2] 刘莹[1] 于秉治[1] 

机构地区：[1]中国医科大学生物化学与分子生物学教研室,辽宁沈阳110001 [2]沈阳市第五人民医院,辽宁沈阳110001

出　　处：《生物工程学报》2016年第9期1204-1211,共8页Chinese Journal of Biotechnology

基　　金：国家自然科学基金(Nos.31501162;81270654;31570819)资助~~

摘　　要：通过研究PKB/Akt与Girdin蛋白之间的关系,目的在于揭示Girdin蛋白在PKB/Akt调控小鼠受精卵微丝聚集中的作用。研究中首先结合软件(http://scansite.mit.edu//)预测了PKB/Akt对Girdin蛋白的磷酸化位点,并制定特定位点磷酸化抗体,通过蛋白免疫印迹检测经PKB/Akt m RNA或PKB/Akt si RNA处理后的小鼠受精卵中Girdin蛋白磷酸化改变情况。同时检测了磷酸化的Girdin蛋白亚细胞定位及同微丝骨架的定位关系。进一步通过显微注射PKB/Akt m RNA再注射Girdin si RNA检测小鼠受精卵微丝骨架的聚集。结果显示经持续激活型PKB/Akt m RNA、野生型PKB/Akt m RNA处理后的小鼠受精卵中p-Girdin-1 417分布位置更加集中在2-细胞分裂沟处。经野生型和持续激活型PKB/Aktm RNA注射组p-Girdin-1 417表达增强,但是并不影响Girdin蛋白的总的表达。说明si RNA介导的PKB/Akt表达敲低非常明显地降低了Girdin蛋白的磷酸化。激光共聚焦研究显示在Akt m RNA和Girdin si RNA共注射组微丝骨架分布明显散乱。本研究结果充分说明Girdin蛋白在小鼠受精卵中受到PKB/Akt的调节,PKB/Akt通过磷酸化Girdin蛋白改变微丝骨架的聚集。The purpose of this study is to reveal the role of Girdin in regulating the aggregation of actin filaments by studying the relationship between PKB/Akt and Girdin. First we used Scansite software（http：//scansite.mit.edu） to predict relevant target sites of PKB/Akt on mouse Girdin. To gain insight into the role of phosphorylation of Girdin by PKB/Akt, we assessed the location of phosphorylated Girdin in fertilized eggs by staining with anti-P-Girdin 1 417 Ab. We detected a distinct increase in the fluorescence signal of F-actin and P-Girdin 1 417 after microinjection of Akt WT and myr-Akt. The addition of myr-Akt induced phosphorylation of Girdin in mouse fertilized eggs. In addition, si RNA-mediated Akt-knockdown blocked phosphorylation of Girdin. The distribution of actin filaments was obviously scattered. These results strongly suggest that PKB/Akt could directly phosphorylate Girdin on Ser 1 417 and promote its function in mouse fertilized eggs.

关 键 词：PKB/AKT Girdin蛋白 F-ACTIN 小鼠受精卵 

分 类 号：Q492[生物学—生理学]