Proteomics Profiling of S-sulfurated Proteins in Acinetobacter baumannii

Brenna J C Walsh; David P Giedroc

doi:10.21769/BioProtoc.4000

Proteomics Profiling of S-sulfurated Proteins in Acinetobacter baumannii

Bio Protoc. 2021 May 5;11(9):e4000. doi: 10.21769/BioProtoc.4000.

Authors

Brenna J C Walsh¹, David P Giedroc^{1

2}

Affiliations

¹ Department of Chemistry, Indiana University, Bloomington, IN 47405 7102, USA.
² Department of Molecular and Cellular Biochemistry, Indiana University, Bloomington, IN 47405, USA.

Abstract

Hydrogen sulfide (H₂S) is emerging as an important modulator in bacterial cytoprotection against the host immune response in infected animals, which may well be attributed to downstream highly oxidized sulfur species, termed reactive sulfur species (RSS), derived from H₂S. One mechanism by which H₂S/RSS may signal in the cell is through proteome S-sulfuration (persulfidation), which is the conversion of protein thiols (-SH) to protein persulfides (-SSH). While several analytical methods have been developed to profile sites of protein persulfidation, few have been applied to bacterial cells. The analytical workflow presented here was recently utilized to profile proteome persulfidation in the major human pathogen Acinetobacter baumannii treated with an exogenous sulfide source, Na₂S. The data obtained using this protocol allow quantitation of the change in persulfidation status of each cysteine in the proteome normalized to the change in protein abundance, thus identifying sites of persulfidation that may constitute regulatory modifications. These can be validated using follow-up biochemical studies.

Keywords: Hydrogen sulfide; Protein persulfidation; Proteomics; Reactive sulfur species.

Abstract

Grants and funding