Document Type
Article
Publication Date
3-2022
Keywords
lysine acetylation; aconitase; TCA cycle; genetic code expansion; deacetylase
Abstract
Aconitase catalyzes the second reaction of the tricarboxylic acid cycle, the reversible conversion of citrate and isocitrate. Escherichia coli has two isoforms of aconitase, AcnA and AcnB. Acetylomic studies have identified acetylation at multiple lysine sites of both E. coli aconitase isozymes, but the impacts of acetylation on aconitases are unknown. In this study, we applied the genetic code expansion approach to produce 14 site-specifically acetylated aconitase variants. Enzyme assays and kinetic analyses showed that acetylation of AcnA K684 decreased the enzyme activity, while acetylation of AcnB K567 increased the enzyme activity. Further in vitro acetylation and deacetylation assays were performed, which indicated that both aconitase isozymes could be acetylated by acetyl-phosphate chemically, and be deacetylated by the CobB deacetylase at most lysine sites. Through this study, we have demonstrated practical applications of genetic code expansion in acetylation studies.
Citation
Araujo, J., Ottinger, S., Venkat, S., Gan, Q., & Fan, C. (2022). Studying Acetylation of Aconitase Isozymes by Genetic Code Expansion. Frontiers in Chemistry, 10, 862483. https://doi.org/10.3389/fchem.2022.862483
Creative Commons License
This work is licensed under a Creative Commons Attribution 4.0 International License.
