Genetically Incorporating Two Distinct Post-translational Modifications into One Protein Simultaneously

ACS Synthetic Biology
Sumana VenkatChenguang Fan

Abstract

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 cotranslationally 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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Citations

Apr 12, 2019·The FEBS Journal·Sumana VenkatChenguang Fan
Jul 11, 2018·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Hao ChenChenguang Fan
Aug 15, 2020·Frontiers in Bioengineering and Biotechnology·Yang WuHao Qi
Nov 9, 2018·Genes·Kyle S HoffmanDieter Söll
Mar 3, 2020·Frontiers in Bioengineering and Biotechnology·Qiuyun XiaoHai Xiong
Oct 30, 2020·Frontiers in Bioengineering and Biotechnology·Zhenling CuiKirill Alexandrov
Dec 16, 2020·Nature Reviews. Genetics·Daniel de la Torre, Jason W Chin
Apr 5, 2021·Current Opinion in Microbiology·Hao ChenChenguang Fan
Jan 16, 2019·ACS Chemical Biology·Hao ChenChenguang Fan
Jun 28, 2019·ACS Chemical Biology·Phillip ZhuRichard B Cooley

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