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大肠杆菌N~α-乙酰基转移酶RimI对人胸腺素α原N末端乙酰化修饰的影响
Effect of N~α-Terminal Acetyltransferase RimI on Human Prothymosin α Acetylation Expressed in Escherichia coli
【摘要】 目的:研究大肠杆菌Nα-乙酰基转移酶RimI对人胸腺素α原(ProTα)N末端乙酰化修饰的影响。方法:在大肠杆菌中共表达ProTα与大肠杆菌Nα-乙酰基转移酶RimI,同时设置只表达ProTα的对照菌,分别提取纯化ProTα,利用HPLC检测其发生乙酰化修饰的程度。结果:ProTα的乙酰化修饰发生在翻译后水平,有无RimI的过表达、诱导时间的长短以及这两个因素之间的交叉效应均对ProTα的乙酰化程度有影响,其F值分别为53.8、89.22及16.28,P值均小于0.0001;无论有无RimI过表达,乙酰化的ProTα所占比例在诱导6 h后逐渐升高(P<0.0001);在过表达Ri-mI的菌株中,乙酰化的ProTα在诱导12 h后占37.4%,而在无RimI过表达的菌株中占64.3%;RimI对ProTα乙酰化的抑制作用从诱导6 h后开始显现(P=0.0007)。结论:在大肠杆菌中,过量的Nα-乙酰基转移酶RimI可抑制人胸腺素α原的乙酰化修饰。
【Abstract】 Objective:To investigate the effect of RimI,one of the Nα-terminal acetyltransferases of E.coli,on human prothymosin α(ProTα) acetylation.Methods:The human ProTα and Nα-terminal acetyltransferase RimI were co-expressed in E.coli DH5α.To observe the level of N-terminal acetylation on human ProTα,ProTα was purified and detected by HPLC.Results:The ProTα was post-translational acetylated.Over-expression of RimI,long term induction and the cross-effect could affect the level of N-terminal acetylation of ProTα(P<0.0001).The increasing N-terminal acetylated ProTα could be observed after inducing 6 h,whether RimI was over-expressed or not(P<0.0001).There were 37.4% N-terminal acetylated ProTα in the presence of overdose RimI,64.3% N-terminal acetylated ProTα without of overdose RimI after inducing 12 h,and we could observe the inhibition after inducing 6 h(P=0.0007).Conclusion:The N-terminal acetylation of ProTα was inhibited when over-expressed Nα-terminal acetyltransferase RimI.
【Key words】 N-terminal acetylation; Nα-terminal acetyltransferase RimI; human prothymosin α; E.coli;
- 【文献出处】 生物技术通讯 ,Letters in Biotechnology , 编辑部邮箱 ,2011年03期
- 【分类号】R378
- 【下载频次】102