Studying Acetylation of Aconitase Isozymes by Genetic Code Expansion

Aconitase catalyzes the second reaction of the tricarboxylic acid cycle, the reversible conversion of citrate and isocitrate. has two isoforms of aconitase, AcnA and AcnB. Acetylomic studies have identified acetylation at multiple lysine sites of both aconitase isozymes, but the impacts of acetylati...

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Bibliographic Details
Published in:Frontiers in chemistry 2022-03, Vol.10, p.862483-862483
Main Authors: Araujo, Jessica, Ottinger, Sara, Venkat, Sumana, Gan, Qinglei, Fan, Chenguang
Format: Article
Language:English
Subjects:
aconitase
Chemistry
deacetylase
genetic code expansion
lysine acetylation
TCA cycle
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Summary:Aconitase catalyzes the second reaction of the tricarboxylic acid cycle, the reversible conversion of citrate and isocitrate. has two isoforms of aconitase, AcnA and AcnB. Acetylomic studies have identified acetylation at multiple lysine sites of both 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 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.
ISSN:2296-2646
2296-2646
DOI:10.3389/fchem.2022.862483