NADH Shuttling Couples Cytosolic Reductive Carboxylation of Glutamine with Glycolysis in Cells with Mitochondrial Dysfunction

Gaude, Edoardo; Schmidt, Christina; Gammage, Payam A.; Dugourd, Aurélien; Blacker, Thomas S.; Chew, Sew Peak; Sáez-Rodríguez, Julio; O’Neill, John S.; Szabadkai, György; Minczuk, Michal; Frezza, Christian

doi:10.1016/j.molcel.2018.01.034

Cited by 180 publications

(193 citation statements)

References 54 publications

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“…4-5) 1,2,6,23,24 . These results are consistent with the earlier experiments 14,15 and with the numerical simulations (Supp. Figs.…”

Section: Resultssupporting

confidence: 93%

“…video 1 and 2). The delicate nature of the presented LDH-GAPDH interactions can explain why it was so difficult to measure that complex 14,15 . The LDH-GAPDH tetramers form a symmetric complex that can support association of multiple LDH and GAPDH molecules in a polymer, which can explain the poor solubility of the complex 14,15 .…”

Section: Resultsmentioning

confidence: 99%

“…11. 14,15 ) Thus, we used extensive numerical simulations 37 in preparation for optimal assay design and data interpretation (Supp. Figs.…”

Section: Resultsmentioning

confidence: 99%

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Substrate Channelingviaa Transient Protein-Protein Complex: The case of D-Glyceraldehyde-3-Phosphate Dehydrogenase and L-Lactate Dehydrogenase

Svedružić

Odorčić

Chang

et al. 2020

Preprint

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Background: D-Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and L-lactate dehydrogenase (LDH) can form a complex that can regulate the major metabolic pathways, however, the exact mechanism remains unknown. We analyzed a possibility of NADHchanneling from GAPDH-NADH complex to LDH isozymes using enzymes from different cells. Results: Enzyme-kinetics and NADH-binding studies showed that LDH can use GAPDH-NADH complex as a substrate. LDH activity with GAPDH-NADH complex was challenged with anti-LDH antibodies to show that the channeled and the diffusive reactions always take place in parallel. The channeling path is dominant only in assays with limiting free-NADH concertation that mimic cytosolic conditions. Analytical ultracentrifugation showed that the channeling does not require a high affinity complex. Molecular dynamics calculations and protein-protein interaction studies showed that LDH and GAPDH can form a leaky channeling complex only at subsaturating NADH concentrations. The interaction sites are conserved between LDH isozymes from heart and muscle, and between GAPDH molecules from rabbit and yeast cells. Positive electric fields between the NAD(H) binding sites on LDH and GAPDH tetramers, showed that NAD(H)-channeling within the LDH-GAPDH complex, can be an extension of NAD(H)channeling between the adjacent subunits in each tetramer. Conclusions: In the case of a transient (GAPDH-NADH)-LDH complex, the relative contribution from the channeled and the diffusive paths depends on the overlap between off-rates for the transient (GAPDH-NADH)-LDH complex and off-rates for the GAPDH-NADH complex. Molecular evolution or metabolic engineering protocols can exploit substrate channeling for metabolic flux control by fine-tuning substrate-binding affinity for the key enzymes in the competing reaction paths. Highlights-Substrate channeling molecular mechanism can regulate energy production and aerobic and anaerobic metabolism in cells -LDH and GAPDH can form a channeling complex only at sub-saturating NADH concentration -Channeled and diffusive paths always compete and take place in parallel -NADH channeling does not require a high-affinity complex -NADH channeling within GAPDH-LDH complex is an extension of NAD(H) channeling within each tetramer -Allosteric modulation of NADH binding affinity in GAPDH tetramer can regulate NAD(H) channeling Introduction D-glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and L-lactate dehydrogenases (LDH) are two NAD(H) dependent dehydrogenases that participate in glycolytic and gluconeogenic pathways 1,2 . Glycolysis and gluconeogenesis are the two major metabolic pathways that provide cells with metabolic precursors and a rapid source of energy. Parallel to glycolysis GAPDH participates in microtubule bundling, DNA replication and repair, apoptosis, the export of nuclear RNA, membrane fusion, and phosphotransferase activity 1,2 . GAPDH is implicated in Huntington's disease 3 , prostate cancer, and viral pathogenesis 1,2 . GAPDH could be a target of nitric oxide 4 and a target of ...

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“…4-5) 1,2,6,23,24 . These results are consistent with the earlier experiments 14,15 and with the numerical simulations (Supp. Figs.…”

Section: Resultssupporting

confidence: 93%

Section: Resultsmentioning

confidence: 99%

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Substrate Channelingviaa Transient Protein-Protein Complex: The case of D-Glyceraldehyde-3-Phosphate Dehydrogenase and L-Lactate Dehydrogenase

Svedružić

Odorčić

Chang

et al. 2020

Preprint

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“…An alternative hypothesis linking mitochondrial dysfunction to glycolysis proposes that reduced oxidative phosphorylation will increase the mitochondrial NADH/NAD + ratio, which then reduces PDH activity. On the other hand, anaerobic glycolysis is activated by a high NADH/NAD + ratio, with LDH consuming the NADH to produce NAD + , contributing to restore the cellular redox state . This process could therefore contribute to the high LDH/PDH ratio observed after CIH exposures.…”

Section: Discussionmentioning

confidence: 99%

“…On the other hand, anaerobic glycolysis is activated by a high NADH/NAD + ratio, with LDH consuming the NADH to produce NAD + , contributing to restore the cellular redox state. 51 This process could therefore contribute to the high LDH/PDH ratio observed after CIH exposures. In this conceptual framework, ER agonists could restore metabolic activity in the brain cortex simply by preventing the mitochondrial dysfunction induced by CIH.…”

Section: Ppt and Dpn Prevent The Metabolic Switch Induced By Cihmentioning

confidence: 99%

Roles of oestradiol receptor alpha and beta against hypertension and brain mitochondrial dysfunction under intermittent hypoxia in female rats

et al. 2019

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Aim Chronic intermittent hypoxia (CIH) induces systemic (hypertension) and central alterations (mitochondrial dysfunction underlying cognitive deficits). We hypothesized that agonists of oestradiol receptors (ER) α and β prevent CIH‐induced hypertension and brain mitochondrial dysfunction. Methods Ovariectomized female rats were implanted with osmotic pumps delivering vehicle (Veh), the ERα agonist propylpyraoletriol (PPT — 30 μg/kg/day) or the ERβ agonist diarylpropionitril (DPN — 100 μg/kg/day). Animals were exposed to CIH (21%‐10% FIO2 — 10 cycles/hour – 8 hours/day – 7 days) or normoxia. Arterial blood pressure was measured after CIH or normoxia exposures. Mitochondrial respiration and H2O2 production were measured in brain cortex with high‐resolution respirometry, as well as activity of complex I and IV of the electron transport chain, citrate synthase, pyruvate, and lactate dehydrogenase (PDH and LDH). Results Propylpyraoletriol but not DPN prevented the rise of arterial pressure induced by CIH. CIH exposures decreased O2 consumption, complex I activity, and increased H2O2 production. CIH had no effect on citrate synthase activity, but decreased PDH activity and increased LDH activity indicating higher anaerobic glycolysis. Propylpyraoletriol and DPN treatments prevented all these alterations. Conclusions We conclude that in OVX female rats, the ERα agonist prevents from CIH‐induced hypertension while both ERα and ERβ agonists prevent the brain mitochondrial dysfunction and metabolic switch induced by CIH. These findings may have implications for menopausal women suffering of sleep apnoea regarding hormonal therapy.

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Mitochondrial DNA mutations in ageing and cancer

2022

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Advancing age is a major risk factor for malignant transformation and the development of cancer. As such, over 50% of neoplasms occur in individuals over the age of 70. The pathologies of both ageing and cancer have been characterized by respective groups of molecular hallmarks, and while some features are divergent between the two pathologies, several are shared. Perturbed mitochondrial function is one such common hallmark, and this observation therefore suggests that mitochondrial alterations may be of significance in age-related cancer development. There is now considerable evidence documenting the accumulation of somatic mitochondrial DNA (mtDNA) mutations in ageing human postmitotic and replicative tissues. Similarly, mutations of the mitochondrial genome have been reported in human cancers for decades. The plethora of functions in which mitochondria partake, such as oxidative phosphorylation, redox balance, apoptosis and numerous biosynthetic pathways, manifests a variety of ways in which alterations in mtDNA may contribute to tumour growth. However, the specific mechanisms by which mtDNA mutations contribute to tumour progression remain elusive and often contradictory. This review aims to consolidate current knowledge and describe future direction within the field.

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NADH Shuttling Couples Cytosolic Reductive Carboxylation of Glutamine with Glycolysis in Cells with Mitochondrial Dysfunction

Cited by 180 publications

References 54 publications

Substrate Channelingviaa Transient Protein-Protein Complex: The case of D-Glyceraldehyde-3-Phosphate Dehydrogenase and L-Lactate Dehydrogenase

Substrate Channelingviaa Transient Protein-Protein Complex: The case of D-Glyceraldehyde-3-Phosphate Dehydrogenase and L-Lactate Dehydrogenase

Roles of oestradiol receptor alpha and beta against hypertension and brain mitochondrial dysfunction under intermittent hypoxia in female rats

Mitochondrial DNA mutations in ageing and cancer

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