Induction of an antioxidant protein of Saccharomyces cerevisiae by O2, Fe3+, or 2-mercaptoethanol.
A soluble 27-kDa protein from Saccharomyces cerevisiae specifically prevents the inactivation of various enzymes caused by a nonenzymatic Fe3+/O2/thiol mixed-function oxidation system but not by mixed-function oxidation systems in which the thiol component is replaced by another electron donor-e.g., ascorbate. In this report, using a 125I-labeled monospecific antibody against the 27-kDa protein, we measured changes in the 27-kDa protector protein in response to changes in oxidative stress and heat shock. With a shift from an anaerobic (95% N2/5% CO2) to a hyperaerobic (95% O2/5% CO2) atmosphere, a 3-fold increase was observed. This increase was prevented by cycloheximide, indicating that the induction requires new protein synthesis. The antioxidant protein synthesis was also significantly enhanced by the addition of either 2-mercaptoethanol or Fe3+ to the growth medium. Radioimmunoassay results also show that the antioxidant protein is an abundant protein, as it constitutes 0.7% of total soluble protein from yeast grown aerobically. Immunoblotting experiments revealed that rat tissues also contain a 27-kDa protein that can be specifically recognized by antibodies against the yeast protein. These results suggest that in vivo induction in yeast of the 27-kDa protein may represent an adaptive response that evolved to protect cells against damage caused by thiol-dependent mixed-function oxidation systems, and the antioxidant protein is conserved in mammalian tissues. A heat shock applied to yeast did not cause any significant increases in the concentration of the 27-kDa protein.
Kim
I H
IH
Laboratory of Biochemistry, National Heart, Lung, and Blood Institute, Bethesda, MD 20892.
Kim
K
K
Rhee
S G
SG
D000332:Aerobiosis; D000693:Anaerobiosis; D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000975:Antioxidants / Q000378:metabolism*; D003513:Cycloheximide / Q000494:pharmacology; D005656:Fungal Proteins / Q000096:biosynthesis* / Q000302:isolation & purification; D006358:Hot Temperature; D007501:Iron / Q000494:pharmacology*; D007700:Kinetics; D008623:Mercaptoethanol / Q000494:pharmacology*; D008970:Molecular Weight; D010100:Oxygen / Q000494:pharmacology*; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000975:Antioxidants; D005656:Fungal Proteins; D008623:Mercaptoethanol; D003513:Cycloheximide; D007501:Iron; D010100:Oxygen