• MeSH
• glycerolfosfátdehydrogenasa MeSH
• ionty MeSH
• kadmium MeSH
• mitochondrie enzymologie   MeSH
• techniky in vitro MeSH

• MeSH
• buněčná membrána MeSH
• glycerolfosfátdehydrogenasa izolace a purifikace   MeSH
• imunoelektroforéza MeSH
• křečci praví MeSH
• kyseliny fosfatidové MeSH
• mitochondrie MeSH
• Check Tag
• křečci praví 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.

• MeSH
• ferrikyanidy metabolismus   MeSH
• glycerolfosfátdehydrogenasa chemie   metabolismus   MeSH
• glycerolfosfáty metabolismus   MeSH
• krysa rodu rattus MeSH
• mitochondrie enzymologie   metabolismus   MeSH
• peroxid vodíku metabolismus   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• savci MeSH
• sukcinátdehydrogenasa chemie   metabolismus   MeSH
• transport elektronů * MeSH
• ubichinon metabolismus   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Prostate cancer is one of the most prominent cancers diagnosed in males. Contrasting with other cancer types, glucose utilization is not increased in prostate carcinoma cells as they employ different metabolic adaptations involving mitochondria as a source of energy and intermediates required for rapid cell growth. In this regard, prostate cancer cells were associated with higher activity of mitochondrial glycerol-3-phosphate dehydrogenase (mGPDH), the key rate limiting component of the glycerophosphate shuttle, which connects mitochondrial and cytosolic processes and plays significant role in cellular bioenergetics. Our research focused on the role of mGPDH biogenesis and regulation in prostate cancer compared to healthy cells. We show that the 42 amino acid presequence is cleaved from N-terminus during mGPDH biogenesis. Only the processed form is part of the mGPDH dimer that is the prominent functional enzyme entity. We demonstrate that mGPDH overexpression enhances the wound healing ability in prostate cancer cells. As mGPDH is at the crossroad of glycolysis, lipogenesis and oxidative metabolism, regulation of its activity by intramitochondrial processing might represent rapid means of cellular metabolic adaptations.

• MeSH
• glycerolfosfátdehydrogenasa metabolismus   MeSH
• HEK293 buňky MeSH
• lidé MeSH
• mitochondrie genetika   metabolismus   MeSH
• nádorové buněčné linie MeSH
• nádory prostaty genetika   metabolismus   MeSH
• transfekce MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

α-Tocopheryl succinate (TOS), a redox-silent analogue of vitamin E, suppresses cell growth in a number of clinical and experimental cancers, inhibits mitochondrial succinate dehydrogenase (SDH) and activates reactive oxygen species (ROS) generation. The aim of this study was to test whether TOS also inhibits glycerol-3-phosphate dehydrogenase (mGPDH), another flavoprotein-dependent enzyme of the mitochondrial respiratory chain because there are differences between electron transfer pathway from SDH and mGPDH to coenzyme Q pool. For our experiments brown adipose tissue mitochondria with high expression of mGPDH were used. Our data showed that inhibition of glycerol-3-phosphate (GP)-dependent oxygen consumption by TOS was more pronounced than the succinate (SUC)-dependent one (50% inhibition was reached at 10 μmol/l TOS vs. 80 μmol/l TOS, respectively). A comparison of the inhibitory effect of TOS on GP-oxidase, GP-cytochrome c oxidoreductase and GP-dehydrogenase activities showed that TOS directly interacts with the dehydrogenase. After TOS application the GP-dependent generation of ROS was highly depressed. It may thus be concluded that TOS-induced inhibition of mGPDH is more pronounced than TOS-induced inhibition of SDH and that the inhibitory effect of TOS for both substrates is exerted at different locations of the particular dehydrogenases. Our data indicate that the inhibition of mGPDH activity could also play a role in TOS-induced growth suppression in neoplastic cells.

• MeSH
• alfa-tokoferol aplikace a dávkování   MeSH
• glycerolfosfátdehydrogenasa antagonisté a inhibitory   biosyntéza   MeSH
• hnědá tuková tkáň enzymologie   MeSH
• karcinogeneze genetika   MeSH
• křečci praví MeSH
• lidé MeSH
• mitochondrie účinky léků   enzymologie   MeSH
• nádory farmakoterapie   enzymologie   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• regulace genové exprese u nádorů účinky léků   MeSH
• spotřeba kyslíku účinky léků   MeSH
• zvířata MeSH
• Check Tag
• křečci praví MeSH
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Mitochondrial glycerol-3-phosphate dehydrogenase (mGPDH) is not included in the traditional textbook schemes of the respiratory chain, reflecting the fact that it is a non-standard, tissue-specific component of mammalian mitochondria. But despite its very simple structure, mGPDH is a very important enzyme of intermediary metabolism and as a component of glycerophosphate shuttle it functions at the crossroads of glycolysis, oxidative phosphorylation and fatty acid metabolism. In this review we summarize the present knowledge on the structure and regulation of mGPDH and discuss its metabolic functions, reactive oxygen species production and tissue and organ specific roles in mammalian mitochondria at physiological and pathological conditions.

• MeSH
• alosterická regulace MeSH
• glycerolfosfátdehydrogenasa genetika   fyziologie   MeSH
• glykolýza MeSH
• lidé MeSH
• mastné kyseliny metabolismus   MeSH
• mitochondrie enzymologie   MeSH
• orgánová specificita MeSH
• oxidativní fosforylace MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• sukcinátdehydrogenasa metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• přehledy MeSH

• MeSH
• boritany farmakologie   chemická syntéza   chemie   MeSH
• glycerolfosfátdehydrogenasa antagonisté a inhibitory   MeSH
• hnědá tuková tkáň MeSH
• inhibitory enzymů farmakologie   chemická syntéza   chemie   MeSH
• křečci praví MeSH
• mitochondrie enzymologie   MeSH
• vztahy mezi strukturou a aktivitou MeSH
• zvířata MeSH
• Check Tag
• křečci praví MeSH
• zvířata MeSH
• Publikační typ
• srovnávací studie MeSH

• MeSH
• glycerolfosfátdehydrogenasa antagonisté a inhibitory   metabolismus   MeSH
• glycerolfosfáty metabolismus   MeSH
• hnědá tuková tkáň enzymologie   ultrastruktura   MeSH
• kinetika MeSH
• křečci praví MeSH
• kyseliny mastné neesterifikované farmakologie   metabolismus   MeSH
• kyseliny olejové farmakologie   metabolismus   MeSH
• mitochondrie enzymologie   účinky léků   MeSH
• zvířata MeSH
• Check Tag
• křečci praví MeSH
• zvířata MeSH

Digitonin solubilizes mitochondrial membrane, breaks the integrity of the respiratory chain and releases two mobile redox-active components: coenzyme Q (CoQ) and cytochrome c (cyt c). In the present study we report the inhibition of glycerol-3-phosphate- and succinate-dependent oxygen consumption rates by digitonin treatment. Our results show that the inhibition of oxygen consumption rates is recovered by the addition of exogenous synthetic analog of CoQ idebenone (hydroxydecyl-ubiquinone; IDB) and cyt c. Glycerol-3-phosphate oxidation rate is recovered to 148 % of control values, whereas succinate-dependent oxidation rate only to 68 %. We find a similar effect on the activities of glycerol-3-phosphate and succinate cytochrome c oxidoreductase. Our results also indicate that succinate-dependent oxidation is less sensitive to digitonin treatment and less activated by IDB in comparison with glycerol-3-phosphate-dependent oxidation. These findings might indicate the different mechanism of the electron transfer from two flavoprotein-dependent dehydrogenases (glycerol-3-phosphate dehydrogenase and succinate dehydrogenase) localized on the outer and inner face of the inner mitochondrial membrane, respectively.

• MeSH
• cytochromy c metabolismus   MeSH
• digitonin farmakologie   MeSH
• glycerolfosfátdehydrogenasa metabolismus   MeSH
• glycerolfosfáty metabolismus   MeSH
• hypertyreóza metabolismus   MeSH
• jaterní mitochondrie metabolismus   účinky léků   MeSH
• kinetika MeSH
• krysa rodu rattus MeSH
• kyselina jantarová metabolismus   MeSH
• mitochondriální membrány metabolismus   účinky léků   MeSH
• modely nemocí na zvířatech MeSH
• obnova funkce MeSH
• oxidace-redukce MeSH
• potkani Wistar MeSH
• spotřeba kyslíku účinky léků   MeSH
• sukcinátcytochrom c oxidoreduktasa metabolismus   MeSH
• ubichinon analogy a deriváty   metabolismus   MeSH
• vztah mezi dávkou a účinkem léčiva MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• MeSH
• glycerolfosfátdehydrogenasa metabolismus   MeSH
• hnědá tuková tkáň enzymologie   metabolismus   MeSH
• křečci praví MeSH
• mitochondrie enzymologie   MeSH
• oxidace-redukce MeSH
• transport elektronů MeSH
• ubichinon analogy a deriváty   analýza   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• křečci praví MeSH
• zvířata MeSH