Changes in disulfide bond content of proteins in a yeast strain lacking major sources of NADPH.
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 beta-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 idp2Deltazwf1Delta 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 idp2Deltazwf1Delta 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 idp2Deltazwf1Delta strain shifted to oleate medium also led to identification of common and unique protein targets of endogenous oxidative stress.
Minard
Karyl I
KI
Department of Biochemistry, University of Texas Health Science Center, San Antonio, TX 78229-3900, USA.
Carroll
Christopher A
CA
Weintraub
Susan T
ST
Mc-Alister-Henn
Lee
L
D019342:Acetic Acid / Q000494:pharmacology; D004220:Disulfides / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional; D005954:Glucosephosphate Dehydrogenase / Q000235:genetics / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007521:Isocitrate Dehydrogenase / Q000235:genetics / Q000378:metabolism; D009249:NADP / Q000737:chemistry / Q000378:metabolism*; D019301:Oleic Acid / Q000494:pharmacology; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D040901:Proteomics; D012441:Saccharomyces cerevisiae / Q000201:enzymology*
D004220:Disulfides; D019301:Oleic Acid; D009249:NADP; D006861:Hydrogen Peroxide; D007521:Isocitrate Dehydrogenase; D005954:Glucosephosphate Dehydrogenase; D019342:Acetic Acid