Genetically Incorporating Two Distinct Post-translational Modifications into One Protein Simultaneously.
Genetically Incorporating Two Distinct Post-translational Modifications into One Protein Simultaneously.
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DOI：
10.1021/acssynbio.7b00408
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发表时间：
2018-02-16
影响因子：
4.7
通讯作者：
Fan C
中科院分区：
文献类型：
--
作者：
Venkat S;Sturges J;Stahman A;Gregory C;Gan Q;Fan C
翻译后修饰在调节多种生物学过程中起着重要作用。为了促进PTM研究，已经利用遗传密码扩展策略将单个PTM（例如乙酰化和磷酸化）共结合到蛋白质的特定位点。然而，最近的研究表明，PTM实际上共同调节蛋白质的功能和结构。因此，将多个不同的PTM同时掺入一种蛋白质中是高度期望的。在这项研究中，我们利用磷酸丝氨酸和乙酰赖氨酸的遗传掺入系统，同时将磷酸化和乙酰化安装到大肠杆菌中的靶蛋白。并利用该系统研究了乙酰化和磷酸化共存对苹果酸脱氢酶的影响，展示了该系统在生物化学研究中的实际应用。此外，我们测试了三种广泛使用的遗传掺入系统的相互正交性，表明将三种不同的PTM同时掺入一种蛋白质的可能性。
Post-translational modifications (PTMs) play important roles in regulating a variety of biological processes. To facilitate PTM studies, the genetic code expansion strategy has been utilized to co-translationally incorporate individual PTMs such as acetylation and phosphorylation into proteins at specific sites. However, recent studies have demonstrated that PTMs actually work together to regulate protein functions and structures. Thus, simultaneous incorporation of multiple distinct PTMs into one protein is highly desirable. In this study, we utilized the genetic incorporation systems of phosphoserine and acetyllysine to install both phosphorylation and acetylation into target proteins simultaneously in Escherichia coli. And we used this system to study the effect of coexisting acetylation and phosphorylation on malate dehydrogenase, demonstrating a practical application of this system in biochemical studies. Furthermore, we tested the mutual orthogonality of three widely-used genetic incorporation systems, indicating the possibility of incorporating three distinct PTMs into one protein simultaneously.
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影响因子：
3.5
作者：
Heinemann IU;Rovner AJ;Aerni HR;Rogulina S;Cheng L;Olds W;Fischer JT;Söll D;Isaacs FJ;Rinehart J
通讯作者：
Rinehart J
影响因子：
14.9
作者：
Fan C;Xiong H;Reynolds NM;Söll D
通讯作者：
Söll D
影响因子：
14.8
作者：
O'Donoghue, Patrick;Ling, Jiqiang;Wang, Yane-Shih;Soell, Dieter
通讯作者：
Soell, Dieter
影响因子：
4.6
作者：
Khoury, George A.;Baliban, Richard C.;Floudas, Christodoulos A.
通讯作者：
Floudas, Christodoulos A.
影响因子：
16.6
作者：
Lee, Sangsik;Oh, Seunghee;Yang, Aerin;Kim, Jihyo;Soell, Dieter;Lee, Daeyoup;Park, Hee-Sung
通讯作者：
Park, Hee-Sung