Chaperones, but not oxidized proteins, are ubiquitinated after oxidative stress.
After oxidative stress, proteins that are oxidatively modified are degraded by the 20S proteasome. However, several studies have documented an enhanced ubiquitination of yet unknown proteins. Because ubiquitination is a prerequisite for degradation by the 26S proteasome in an ATP-dependent manner this raises the question whether these proteins are also oxidized and, if not, what proteins need to be ubiquitinated and degraded after oxidative conditions. By determination of oxidized and ubiquitinated proteins we demonstrate here that most oxidized proteins are not preferentially ubiquitinated. However, we were able to confirm an increase in ubiquitinated proteins 16 h after oxidative stress. Therefore, we isolated ubiquitinated proteins from hydrogen peroxide-treated cells, as well as from control cells and cells treated with lactacystin, an irreversible proteasome inhibitor, and identified some of these proteins by MALDI tandem mass spectrometry. As a result we obtained 24 different proteins that can be categorized into the following groups: chaperones, energy metabolism, cytoskeleton/intermediate filaments, and protein translation/ribosome biogenesis. The special set of identified, ubiquitinated proteins confirms the thesis that ubiquitination upon oxidative stress is not a random process to degrade the mass of oxidized proteins, but concerns a special group of functional proteins.
Department of Nutritional Toxicology, Institute of Nutrition, Friedrich-Schiller University Jena, 07743 Jena, Germany.
Marc
M
Kästle
Sandra
S
Reeg
Adelina
A
Rogowska-Wrzesinska
Tilman
T
Grune
D000111:Acetylcysteine / Q000031:analogs & derivatives / Q000494:pharmacology; D045744:Cell Line, Tumor; D003599:Cytoskeleton / Q000378:metabolism; D004734:Energy Metabolism / Q000502:physiology; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D018832:Molecular Chaperones / Q000737:chemistry / Q000378:metabolism*; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D046988:Proteasome Endopeptidase Complex / Q000187:drug effects / Q000378:metabolism; D061988:Proteasome Inhibitors / Q000494:pharmacology; D014176:Protein Biosynthesis; D011506:Proteins / Q000737:chemistry / Q000378:metabolism*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D025801:Ubiquitin / Q000378:metabolism*; D057149:Ubiquitinated Proteins / Q000737:chemistry / Q000378:metabolism*; D054875:Ubiquitination
D018832:Molecular Chaperones; D061988:Proteasome Inhibitors; D011506:Proteins; D025801:Ubiquitin; D057149:Ubiquitinated Proteins; C067713:lactacystin; D006861:Hydrogen Peroxide; D046988:Proteasome Endopeptidase Complex; D000111:Acetylcysteine