Protocol for using organic persulfides to measure the chemical reactivity of persulfide sensors

Joseph N Fakhoury; Daiana A Capdevila; David P Giedroc

doi:10.1016/j.xpro.2022.101424

Protocol for using organic persulfides to measure the chemical reactivity of persulfide sensors

STAR Protoc. 2022 May 25;3(2):101424. doi: 10.1016/j.xpro.2022.101424. eCollection 2022 Jun 17.

Authors

Joseph N Fakhoury¹, Daiana A Capdevila¹, David P Giedroc¹

Affiliation

¹ Department of Chemistry, Indiana University, 800 E. Kirkwood Avenue, Bloomington, IN 47405-7102, USA.

Abstract

Hydrogen sulfide (H₂S) and downstream reactive sulfur species (RSS), including organic persulfides, protect bacterial cells against diverse oxidative stressors. Specialized dithiol-based transcriptional repressors sense persulfides directly to control cellular H₂S/RSS and avoid toxicity. Here, we present a protocol to quantify the kinetics of chemical reactivity of cysteines in two bacterial persulfide sensors toward cysteine persulfide and glutathione persulfide, with a LC-ESI-MS analysis that results in a kinetic model. This protocol has potential applications to other cysteine-containing proteins and oxidants. For complete details on the use and execution of this protocol, please refer to Fakhoury et al. (2021) and Capdevila et al. (2021).

Keywords: Mass Spectrometry; Protein Biochemistry.

Publication types

Research Support, Non-U.S. Gov't
Research Support, N.I.H., Extramural

MeSH terms

Chromatography, Liquid
Hydrogen Sulfide* / metabolism
Oxidation-Reduction
Sulfides* / metabolism

Substances

Sulfides
persulfides
Hydrogen Sulfide

Grants and funding

R35 GM118157/GM/NIGMS NIH HHS/United States