Bacillus subtilis YtpP and thioredoxin A are new players in the CoA-mediated defense mechanism against cellular stress

ABSTRACT: Coenzyme A is an important cellular metabolite critical for metabolic processes and the regulation of gene expression. The recent discovery of its antioxidant role has highlighted its protective function that leads to the formation of a mixed-disulfide bond with protein cysteines, termed protein CoAlation. To date, more than 2,000 CoAlated bacterial and mammalian proteins have been identified in cellular response to oxidative stress with the majority being involved in metabolic pathways (60%). In vitro and cellular studies have shown that protein CoAlation is a widespread post-translational modification, which modulates the activity and conformation of the modified proteins. The induction of protein CoAlation by oxidative stress was found to be rapidly reversed after the removal of oxidizing agents from the medium of cultured cells. In this study, we developed an ELISA-based deCoAlation assay to detect deCoAlation activity from B. subtilis and B. megaterium lysates. We then used a combination of ELISA-based assay and purification strategies to show that deCoAlation is an enzyme-driven mechanism. Using mass-spectrometry and deCoAlation assays (anti-CoA WB), we identified B. subtilis YtpP (thioredoxin-like protein) and thioredoxin A (TrxA) as enzymes that can remove CoA from different substrates. With mutagenesis studies, we identified YtpP and TrxA catalytic cysteine residues and proposed a possible deCoAlation mechanism for MsrA and PRDX5, which results in the release of both CoA and the reduced form of MsrA or PRDX5. Overall, this paper reveals deCoAlation activity of YtpP and TrxA, and opens doors to future studies on the CoA-mediated redox regulation of CoAlated proteins under various cellular stress conditions.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Bacillus Subtilis Subsp. Subtilis Str. L170

SUBMITTER: Mark Skehel  

LAB HEAD: Ivan Gout

PROVIDER: PXD041347 | Pride | 2023-05-10

REPOSITORIES: Pride