Oxidative stress studies in yeast with a frataxin mutant: a proteomics perspective.
Cellular response of wild-type Saccharomyces cerevisiae and the Delta yfh1 mutant to oxidative stress (OS) was examined by stressing cells through the addition of H(2)O(2) to the growth medium. The Delta yfh1 mutant is unusual in that it accumulates iron in it is mitochondria. Wild-type growth was immediately arrested and recovered in 2 h following H(2)O(2) treatment. No change in viability was observed. Growth of the mutant, on the other hand, was similar to wild-type yeast for 4 h but then rapidly declined, eventually reaching zero. Levels of carbonyl groups and reactive oxygen species (ROS) reached their maximum at 3 h following exposure. The impact of OS on protein function was also evaluated by proteomic techniques targeting protein carbonylation. Oxidized proteins were selected by affinity chromatography, and following trypsin digestion, peptide fragments were identified by RPLC-MS/MS. A total of 53 proteins were identified in both wild-type and mutant cells, respectively.
Kim
Jin-Hee
JH
Department of Chemistry, Purdue University, West Lafayette, Idiana 47907, USA.
Sedlak
Miroslav
M
Gao
Qiang
Q
Riley
Catherine P
CP
Regnier
Fred E
FE
Adamec
Jiri
J
D002846:Chromatography, Affinity; D003470:Culture Media; D033862:Iron-Binding Proteins / Q000235:genetics*; D009154:Mutation; D018384:Oxidative Stress; D040901:Proteomics; D017382:Reactive Oxygen Species / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D000096923:Frataxin
D003470:Culture Media; D033862:Iron-Binding Proteins; D017382:Reactive Oxygen Species; D000096923:Frataxin