Design, Validation, and Application of an Enzyme-Coupled Hydrogen Sulfide Detection Assay.

ABSTRACT: Hydrogen sulfide (H₂S) is a key metabolite in biosynthesis and is increasingly being recognized as an essential gasotransmitter. Owing to its diffusible and reactive nature, H₂S can be difficult to quantify, particularly in situ. Although several detection schemes are available, they have drawbacks. In efforts to quantify sulfide release in the cross-linking reaction of the flagellar protein FlgE, we developed an enzyme-coupled sulfide detection assay using the Escherichia coli O-acetylserine sulfhydrylase enzyme CysM. Conversion of HS^- to l-cysteine via CysM followed by derivatization with the thiol-specific fluorescent dye 7-diethylamino-3-(4-maleimidophenyl)-4-methylcoumarin enables for facile detection and quantification of H₂S by fluorescent

SUBMITTER: Lynch MJ 

PROVIDER: S-EPMC6526949 | biostudies-literature | 2019 Feb

REPOSITORIES: biostudies-literature