Glutathione peroxidase 3 of Saccharomyces cerevisiae regulates the activity of methionine sulfoxide reductase in a redox state-dependent way.
Glutathione peroxidase (Gpx) is one of the most important anti-oxidant enzymes in yeast. Gpx3 is a ubiquitously expressed isoform that modulates the activities of redox-sensitive thiol proteins, particularly those involved in signal transduction pathways and protein translocation. In order to search for the interaction partners of Gpx3, we carried out immunoprecipitation/2-dimensional gel electrophoresis (IP-2DE), MALDI-TOF mass spectrometry, and a pull down assay. We found that Mxr1, a peptide methionine sulfoxide reductase, interacts with Gpx3. By reducing methionine sulfoxide to methionine, Mxr1 reverses the inactivation of proteins caused by the oxidation of critical methionine residues. Gpx3 can interact with Mxr1 through the formation of an intermolecular disulfide bond. When oxidative stress is induced by H(2)O(2), this interaction is compromised and the free Mxr1 then repairs the oxidized proteins. Our findings imply that this interaction links redox sensing machinery of Gpx3 to protein repair activity of Mxr1. Based on these results, we propose that Gpx3 functions as a redox-dependent exquisite regulator of the protein repair activity of Mxr1.
Kho
Chang Won
CW
Proteome Research Laboratory, Korea Research Institute of Bioscience and Biotechnology, Daejeon 305-333, Republic of Korea.
Lee
Phil Young
PY
Bae
Kwang-Hee
KH
Cho
Sayeon
S
Lee
Zee-Won
ZW
Park
Byoung Chul
BC
Kang
Seongman
S
Lee
Do Hee
DH
Park
Sung Goo
SG
D004789:Enzyme Activation; D005979:Glutathione Peroxidase / Q000235:genetics / Q000378:metabolism*; D051150:Methionine Sulfoxide Reductases; D009154:Mutation; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D010088:Oxidoreductases / Q000378:metabolism*; D017382:Reactive Oxygen Species / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000254:growth & development; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*
D017382:Reactive Oxygen Species; D011994:Recombinant Proteins; D029701:Saccharomyces cerevisiae Proteins; D010088:Oxidoreductases; D005979:Glutathione Peroxidase; C494438:HYR1 protein, S cerevisiae; D051150:Methionine Sulfoxide Reductases; C027219:methionine sulfoxide reductase