Mapping of sulfur metabolic pathway by LC Orbitrap mass spectrometry

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Journal titleAnalytica Chimica Acta
Pages129136; # of pages: 8
SubjectThiol; Yeast; LTQ-Orbitrap mass spectrometry; Oxidative stress; Metabolic study
AbstractFor the first time a liquid chromatography method with high resolution mass spectrometric detection has been developed for the simultaneous determination all key metabolites of the sulfur pathway in yeast, including all thiolic (cysteine (Cys), homocysteine (HCys), glutathione (GSH), cysteinyl-glycine (Cys-Gly), γ-glutamyl-cysteine (Glu-Cys)) and non-thiolic compounds (methionine (Met), s-adenosyl-methionine (AdoMet), s-adenosyl-homocysteine (AdoHcy), and cystathionine (Cysta)). The developed assay also permits the speciation and selective determination of reduced, oxidized and protein bound fractions of all of the five thiols. Iodoacetic acid (IAA) was chosen as the derivatizing reagent. Thiols were extracted from sub-mg quantities of yeast using hot 75% ethanol. The detection limits were in the range of 1–12 nmol L−¹ for standard solution (high femotomole, absolute), except AdoMet (116 nmol L−¹), which was unstable. In freshly harvested yeast, most of the thiols were in the reduced forms and low levels of protein-bound GSH and Glu-Cys were found. In a selenium enriched yeast, the thiols were mainly in the oxidized forms, and a significant amount of protein-bound Cys, HCys, GSH, Cys-Gly and Glu-Cys were found. The method was also applied to the metabolic study of the adaptive response of Saccharomyces cerevisiae to hydrogen peroxide, cadmium, and arsenite, and the change in concentration of thiols in the sulfur pathway was monitored over a period of 4 h.
Publication date
AffiliationMeasurement Science and Standards; National Research Council Canada
Peer reviewedYes
NPARC number21268610
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Record identifier4900d65e-ec86-4371-ae3b-ebe7be0593bf
Record created2013-10-28
Record modified2016-05-09
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