Increasing Inhibition of the Rat Brain 2-Oxoglutarate Dehydrogenase Decreases Glutathione Redox State, Elevating Anxiety and Perturbing Stress Adaptation

Artiukhov, Artem V.; Граф, А. В.; Kazantsev, A. V.; Boyko, Alexandra; Aleshin, Vasily A.; Ksenofontov, Alexander L.; Bunik, Victoria I.

doi:10.3390/ph15020182

Cited by 9 publications

(26 citation statements)

References 110 publications

(140 reference statements)

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“…However, neither cystine, nor citrulline levels are significantly reduced by the TMAP treatment of SCI animals, suggesting another mechanism of the glutathione drop after SCI. Among these mechanisms, OGDHC dysfunction is a factor that may perturb the redox status of the brain glutathione buffer ( 55 ). Yet in our experimental settings, the TMAP treatment is not significant for the OGDHC activity, which responds to the (patho)physiological state only ( Figure 5C , condition significance).…”

Section: Resultsmentioning

confidence: 99%

Delayed Impact of 2-Oxoadipate Dehydrogenase Inhibition on the Rat Brain Metabolism Is Linked to Protein Glutarylation

et al. 2022

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BackgroundThe DHTKD1-encoded 2-oxoadipate dehydrogenase (OADH) oxidizes 2-oxoadipate—a common intermediate of the lysine and tryptophan catabolism. The mostly low and cell-specific flux through these pathways, and similar activities of OADH and ubiquitously expressed 2-oxoglutarate dehydrogenase (OGDH), agree with often asymptomatic phenotypes of heterozygous mutations in the DHTKD1 gene. Nevertheless, OADH/DHTKD1 are linked to impaired insulin sensitivity, cardiovascular disease risks, and Charcot-Marie-Tooth neuropathy. We hypothesize that systemic significance of OADH relies on its generation of glutaryl residues for protein glutarylation. Using pharmacological inhibition of OADH and the animal model of spinal cord injury (SCI), we explore this hypothesis.MethodsThe weight-drop model of SCI, a single intranasal administration of an OADH-directed inhibitor trimethyl adipoyl phosphonate (TMAP), and quantification of the associated metabolic changes in the rat brain employ established methods.ResultsThe TMAP-induced metabolic changes in the brain of the control, laminectomized (LE) and SCI rats are long-term and (patho)physiology-dependent. Increased glutarylation of the brain proteins, proportional to OADH expression in the control and LE rats, represents a long-term consequence of the OADH inhibition. The proportionality suggests autoglutarylation of OADH, supported by our mass-spectrometric identification of glutarylated K155 and K818 in recombinant human OADH. In SCI rats, TMAP increases glutarylation of the brain proteins more than OADH expression, inducing a strong perturbation in the brain glutathione metabolism. The redox metabolism is not perturbed by TMAP in LE animals, where the inhibition of OADH increases expression of deglutarylase sirtuin 5. The results reveal the glutarylation-imposed control of the brain glutathione metabolism. Glutarylation of the ODP2 subunit of pyruvate dehydrogenase complex at K451 is detected in the rat brain, linking the OADH function to the brain glucose oxidation essential for the redox state. Short-term inhibition of OADH by TMAP administration manifests in increased levels of tryptophan and decreased levels of sirtuins 5 and 3 in the brain.ConclusionPharmacological inhibition of OADH affects acylation system of the brain, causing long-term, (patho)physiology-dependent changes in the expression of OADH and sirtuin 5, protein glutarylation and glutathione metabolism. The identified glutarylation of ODP2 subunit of pyruvate dehydrogenase complex provides a molecular mechanism of the OADH association with diabetes.

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Section: Resultsmentioning

confidence: 99%

Delayed Impact of 2-Oxoadipate Dehydrogenase Inhibition on the Rat Brain Metabolism Is Linked to Protein Glutarylation

et al. 2022

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“…Quantification of low-molecular weight metabolites in the methanol/acetic acid extracts of rat cerebral cortex was performed as described previously ( Artiukhov et al, 2022 ). The extracts were prepared according to Ksenofontov et al (2017 ): the brain tissue (0.5 g) was homogenized in 4 ml ice-cold methanol, with the resulting homogenate diluted with 0.2% acetic acid at a 1:1.5 ratio.…”

Section: Methodsmentioning

confidence: 99%

“…The chromatograms were processed using MultiChrom for Windows software (Ampersand Ltd., Moscow, Russia). Levels of NAD + and oxidized glutathione were measured with previously described fluorometric assays ( Artiukhov et al, 2022 ) in a microplate format using a CLARIOstar Plus microplate reader (BMG Labtech, Ortenberg, Germany).…”

Section: Methodsmentioning

confidence: 99%

“…Enzyme activities were measured spectrophotometrically in cerebral cortex homogenates as described in previous studies ( Araujo et al, 2013 ; Tsepkova et al, 2017 ; Artiukhov et al, 2022 ). The brain tissue (0.5 g) was homogenized in 1.25 ml of 50 mM MOPS, pH = 7.0, containing 2.7 mM EDTA, 20% glycerol, and protease inhibitors (0.2 mM AEBSF, 0.16 µM aprotinin, 3.33 µM bestatin, 3 µM E-64, 2 µM leupeptin, and 1.4 µM pepstatin A) using ULTRA-TURRAX ® T-10 basic disperser (IKA, Staufen, Germany).…”

Section: Methodsmentioning

confidence: 99%

“…The activities of glutamate dehydrogenase, malate dehydrogenase, NADP + -dependent malic enzyme, OGDHC, and extramitochondrial OADHC were detected using absorbance of NAD(P)H at 340 nm ( Tsepkova et al, 2017 ). PDHC activity was detected by absorbance of the iodonitrotetrazolium-formazan product at 500 nm ( Artiukhov et al, 2022 ). Glutamine synthetase activity was assayed by absorbance of the γ-glutamilhydoxamate-Fe 3+ complex at 540 nm after 15 min of the start of the reaction ( Levintow, 1954 ).…”

Section: Methodsmentioning

confidence: 99%

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Administration of Phosphonate Inhibitors of Dehydrogenases of 2-Oxoglutarate and 2-Oxoadipate to Rats Elicits Target-Specific Metabolic and Physiological Responses

et al. 2022

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In vitro and in cell cultures, succinyl phosphonate (SP) and adipoyl phosphonate (AP) selectively target dehydrogenases of 2-oxoglutarate (OGDH, encoded by OGDH/OGDHL) and 2-oxoadipate (OADH, encoded by DHTKD1), respectively. To assess the selectivity in animals, the effects of SP, AP, and their membrane-penetrating triethyl esters (TESP and TEAP) on the rat brain metabolism and animal physiology are compared. Opposite effects of the OGDH and OADH inhibitors on activities of OGDH, malate dehydrogenase, glutamine synthetase, and levels of glutamate, lysine, citrulline, and carnosine are shown to result in distinct physiological responses. ECG is changed by AP/TEAP, whereas anxiety is increased by SP/TESP. The potential role of the ester moiety in the uncharged precursors of the 2-oxo acid dehydrogenase inhibitors is estimated. TMAP is shown to be less efficient than TEAP, in agreement with lower lipophilicity of TMAP vs. TEAP. Non-monotonous metabolic and physiological impacts of increasing OADH inhibition are revealed. Compared to the non-treated animals, strong inhibition of OADH decreases levels of tryptophan and beta-aminoisobutyrate and activities of malate dehydrogenase and pyruvate dehydrogenase, increasing the R–R interval of ECG. Thus, both metabolic and physiological actions of the OADH-directed inhibitors AP/TEAP are different from those of the OGDH-directed inhibitors SP/TESP, with the ethyl ester being more efficient than methyl ester.

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Phosphonate Inhibitors of Pyruvate Dehydrogenase Perturb Homeostasis of Amino Acids and Protein Succinylation in the Brain

Artiukhov¹,

Aleshin²,

Karlina³

et al. 2022

Preprint

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Mitochondrial pyruvate dehydrogenase complex (PDHC) is essential for the brain glucose and neurotransmitter metabolism, dysregulated in many pathologies. Using specific inhibitors of PDHC in vivo, we determine biochemical and physiological responses to PDHC dysfunction. Dose dependence of the responses to membrane-permeable dimethyl acetylphosphonate (Ac-PMe2) is non-monotonous. Primary decreases in glutathione and its redox potential, methionine and ethanolamine are alleviated with increasing PDHC inhibition, the alleviation accompanied by physiological changes. Comparison of 39 brain biochemical parameters after administration of four phosphinate and phosphonate analogs of pyruvate at a fixed dose of 0.1 mmol/kg reveals no primary, but the secondary changes, such as activation of 2-oxoglutarate dehydrogenase complex (OGDHC) and decreased levels of glutamate, isoleucine and leucine. The accompanying decreases in RMSSD of ECG and freezing time are most pronounced after administration of me-thyl acetylphosphinate and dimethyl acetylphosphonate. The levels of PDHA1 expression and phosphorylation, sirtuin 3 and total protein acetylation are not significantly changed by the PDHC inhibitors that affect the brain protein succinylation and glutarylation. Thus, decreased production of the tricarboxylic acid cycle substrate acetyl-CoA by inhibited PDHC is compen-sated by increased degradation of amino acids through the cycle with activated OGDHC, increas-ing total protein succinylation and decreasing anxiety indicators.

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Increasing Inhibition of the Rat Brain 2-Oxoglutarate Dehydrogenase Decreases Glutathione Redox State, Elevating Anxiety and Perturbing Stress Adaptation

Cited by 9 publications

References 110 publications

Delayed Impact of 2-Oxoadipate Dehydrogenase Inhibition on the Rat Brain Metabolism Is Linked to Protein Glutarylation

Delayed Impact of 2-Oxoadipate Dehydrogenase Inhibition on the Rat Brain Metabolism Is Linked to Protein Glutarylation

Administration of Phosphonate Inhibitors of Dehydrogenases of 2-Oxoglutarate and 2-Oxoadipate to Rats Elicits Target-Specific Metabolic and Physiological Responses

Phosphonate Inhibitors of Pyruvate Dehydrogenase Perturb Homeostasis of Amino Acids and Protein Succinylation in the Brain

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