ROS generation and multiple forms of mammalian mitochondrial glycerol-3-phosphate dehydrogenase
Language English Country Netherlands Media print-electronic
Document type Journal Article, Research Support, Non-U.S. Gov't
PubMed
23999537
DOI
10.1016/j.bbabio.2013.08.007
PII: S0005-2728(13)00144-8
Knihovny.cz E-resources
- Keywords
- 2,6-dichlorophenolindophenol, 2-n-heptyl-4-hydroxyquinoline N-oxide, AA, BAT, CoQ, DCPIP, E.C. 1.1.5.3, FeCN, GP, HAR, HQNO, HRP, In-gel ROS detection, MXT, Mitochondrial glycerol-3-phosphate dehydrogenase, OXPHOS, ROS, ROS production, SDH, Supercomplex, antimycin A, brown adipose tissue, coenzyme Q, ferricyanide, potassium hexacyanoferrate(III), hexaammineruthenium(III) chloride, horseradish peroxidase, mGPDH, mitochondrial FAD-dependent glycerol-3-phosphate dehydrogenase, myxothiazol, oxidative phosphorylation, reactive oxygen species, sn-glycerol-3-phosphate, succinate dehydrogenase,
- MeSH
- Ferricyanides metabolism MeSH
- Glycerolphosphate Dehydrogenase chemistry metabolism MeSH
- Glycerophosphates metabolism MeSH
- Rats MeSH
- Mitochondria enzymology metabolism MeSH
- Hydrogen Peroxide metabolism MeSH
- Reactive Oxygen Species metabolism MeSH
- Mammals MeSH
- Succinate Dehydrogenase chemistry metabolism MeSH
- Electron Transport * MeSH
- Ubiquinone metabolism MeSH
- Animals MeSH
- Check Tag
- Rats MeSH
- Animals MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
- Names of Substances
- Ferricyanides MeSH
- Glycerolphosphate Dehydrogenase MeSH
- Glycerophosphates MeSH
- Hydrogen Peroxide MeSH
- Reactive Oxygen Species MeSH
- Succinate Dehydrogenase MeSH
- Ubiquinone MeSH
Overproduction of reactive oxygen species (ROS) has been implicated in a range of pathologies. Mitochondrial flavin dehydrogenases glycerol-3-phosphate dehydrogenase (mGPDH) and succinate dehydrogenase (SDH) represent important ROS source, but the mechanism of electron leak is still poorly understood. To investigate the ROS production by the isolated dehydrogenases, we used brown adipose tissue mitochondria solubilized by digitonin as a model. Enzyme activity measurements and hydrogen peroxide production studies by Amplex Red fluorescence, and luminol luminescence in combination with oxygraphy revealed flavin as the most likely source of electron leak in SDH under in vivo conditions, while we propose coenzyme Q as the site of ROS production in the case of mGPDH. Distinct mechanism of ROS production by the two dehydrogenases is also apparent from induction of ROS generation by ferricyanide which is unique for mGPDH. Furthermore, using native electrophoretic systems, we demonstrated that mGPDH associates into homooligomers as well as high molecular weight supercomplexes, which represent native forms of mGPDH in the membrane. By this approach, we also directly demonstrated that isolated mGPDH itself as well as its supramolecular assemblies are all capable of ROS production.
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