Changes in disulfide bond content of proteins in a yeast strain lacking major sources of NADPH

Karyl I. Minard, Christopher A. Carroll, Susan E Weintraub, Lee Mc-Alister-Henn

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

A yeast mutant lacking the two major cytosolic sources of NADPH, glucose-6-phosphate dehydrogenase (Zwf1p) and NADP+-specific isocitrate dehydrogenase (Idp2p), has been demonstrated to lose viability when shifted to medium with acetate or oleate as the carbon source. This loss in viability was found to correlate with an accumulation of endogenous oxidative by-products of respiration and peroxisomal β-oxidation. To assess effects on cellular protein of endogenous versus exogenous oxidative stress, a proteomics approach was used to compare disulfide bond-containing proteins in the idp2Δzwf1Δ strain following shifts to acetate and oleate media with those in the parental strain following similar shifts to media containing hydrogen peroxide. Among prominent disulfide bond-containing proteins were several with known antioxidant functions. These and several other proteins were detected as multiple electrophoretic isoforms, with some isoforms containing disulfide bonds under all conditions and other isoforms exhibiting a redox-sensitive content of disulfide bonds, i.e., in the idp2Δzwf1Δ strain and in the hydrogen peroxide-challenged parental strain. The disulfide bond content of some isoforms of these proteins was also elevated in the parental strain grown on glucose, possibly suggesting a redirection of NADPH reducing equivalents to support rapid growth. Further examination of protein carbonylation in the idp2Δzwf1Δ strain shifted to oleate medium also led to identification of common and unique protein targets of endogenous oxidative stress.

Original languageEnglish (US)
Pages (from-to)106-117
Number of pages12
JournalFree Radical Biology and Medicine
Volume42
Issue number1
DOIs
StatePublished - Jan 1 2007

Fingerprint

NADP
Disulfides
Yeast
Yeasts
Protein Isoforms
Oleic Acid
Proteins
Oxidative stress
Hydrogen Peroxide
Acetates
Oxidative Stress
Protein Carbonylation
Glucosephosphate Dehydrogenase
Carbonylation
Isocitrate Dehydrogenase
Proteomics
Oxidation-Reduction
Respiration
Carbon
Antioxidants

Keywords

  • Carbonylation
  • Disulfide bonds
  • Glucose-6-phosphate dehydrogenase
  • Isocitrate dehydrogenase
  • NADPH
  • Reactive thiols

ASJC Scopus subject areas

  • Medicine(all)
  • Toxicology
  • Clinical Biochemistry

Cite this

Changes in disulfide bond content of proteins in a yeast strain lacking major sources of NADPH. / Minard, Karyl I.; Carroll, Christopher A.; Weintraub, Susan E; Mc-Alister-Henn, Lee.

In: Free Radical Biology and Medicine, Vol. 42, No. 1, 01.01.2007, p. 106-117.

Research output: Contribution to journalArticle

Minard, Karyl I. ; Carroll, Christopher A. ; Weintraub, Susan E ; Mc-Alister-Henn, Lee. / Changes in disulfide bond content of proteins in a yeast strain lacking major sources of NADPH. In: Free Radical Biology and Medicine. 2007 ; Vol. 42, No. 1. pp. 106-117.
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