Isolation and Characterization of a Mammalian Hydroxyacid-Oxoacid Transhydrogenase

Kaufman, Elaine E.; Nelson, Thomas; Fales, Henry M.

doi:10.1016/b978-0-12-584552-6.50040-9

Cited by 32 publications

(59 citation statements)

References 13 publications

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“…The conversion of D-2-HG to 2-ketoglutarate is catalyzed by D-2-HG dehydrogenase located in the mitochondria (15 ). A second mitochondrial enzyme, hydroxyacid-oxoacid transhydrogenase, also converts D-2-HG to 2-ketoglutarate, with succinic semialdehyde converted to ␥-hydroxybutyrate in the same reaction (16 ). In humans, the existence of the latter enzyme has not been reported.…”

Section: Discussionmentioning

confidence: 99%

Disease-related Metabolites in Culture Medium of Fibroblasts from Patients with d-2-Hydroxyglutaric Aciduria, l-2-Hydroxyglutaric Aciduria, and Combined d/l-2-Hydroxyglutaric Aciduria

et al. 2003

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Background: d-2-Hydroxyglutaric aciduria (D-2-HGA), l-2-hydroxyglutaric aciduria (L-2-HGA), and the combined d/l-2-hydroxyglutaric aciduria (D/L-2-HGA) are poorly understood organic acidurias. To investigate the usefulness of cultured human skin fibroblasts for both diagnostic and research purposes, we measured disease-related metabolites in the cell culture medium. Methods: We measured d-2-hydroxyglutarate (D-2-HG), l-2-hydroxyglutarate (L-2-HG), succinate, 2-ketoglutarate, and citrate in fibroblast cell medium by stable-isotope-dilution gas chromatography–mass spectrometry and glutamine, glutamic acid, and lysine with an amino acid analyzer. We used six cell lines from patients with D-2-HGA, two from patients with L-2-HGA, three from patients with D/L-2-HGA, and seven control cell lines. Culture medium was analyzed after a 96-h incubation period. Results: Culture media from cell lines from D-2-HGA patients contained D-2-HG at concentrations 5- to 30-fold higher than media from controls, whereas the concentration of L-2-HG in media was not increased. Media from L-2-HGA cell lines showed a fivefold increase in L-2-HG compared with controls. Media containing fibroblasts from D/L-2-HGA patients contained moderately increased amounts of both D-2-HG and L-2-HG. For all cell lines, succinate concentrations in the blank medium were higher than after 96 h of incubation with the exception of two of three D/L-2-HGA cell lines. Media of D-2-HGA cell lines had 2-ketoglutarate concentrations that were 40% of that for controls. Glutamic acid concentrations in media of these cell lines were 60% lower than in controls. Conclusions: Cell culture media from fibroblasts from patients with D-2-HGA, L-2-HGA, or D/L-2-HGA contain increased amounts the corresponding 2-HGs, demonstrating the suitability of fibroblasts for both diagnosis of and research concerning 2-HGAs.

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Section: Discussionmentioning

confidence: 99%

Disease-related Metabolites in Culture Medium of Fibroblasts from Patients with d-2-Hydroxyglutaric Aciduria, l-2-Hydroxyglutaric Aciduria, and Combined d/l-2-Hydroxyglutaric Aciduria

et al. 2003

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“…Another candidate source of intracellular 2HG is the poorly studied mitochondrial enzyme ADHFE1 (28,48). This enzyme did not seem to be elevated in breast tumors based on gene expression data.…”

Section: Discussionmentioning

confidence: 99%

MYC-driven accumulation of 2-hydroxyglutarate is associated with breast cancer prognosis

et al. 2013

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Metabolic profiling of cancer cells has recently been established as a promising tool for the development of therapies and identification of cancer biomarkers.Here we characterized the metabolomic profile of human breast tumors and uncovered intrinsic metabolite signatures in these tumors using an untargeted discovery approach and validation of key metabolites. The oncometabolite 2-hydroxyglutarate (2HG) accumulated at high levels in a subset of tumors and human breast cancer cell lines. We discovered an association between increased 2HG levels and MYC pathway activation in breast cancer, and further corroborated this relationship using MYC overexpression and knockdown in human mammary epithelial and breast cancer cells. Further analyses revealed globally increased DNA methylation in 2HG-high tumors and identified a tumor subtype with high tissue 2HG and a distinct DNA methylation pattern that was associated with poor prognosis and occurred with higher frequency in African-American patients. Tumors of this subtype had a stem cell-like transcriptional signature and tended to overexpress glutaminase, suggestive of a functional relationship between glutamine and 2HG metabolism in breast cancer. Accordingly, 13C-labeled glutamine was incorporated into 2HG in cells with aberrant 2HG accumulation, whereas pharmacologic and siRNAmediated glutaminase inhibition reduced 2HG levels. Our findings implicate 2HG as a candidate breast cancer oncometabolite associated with MYC activation and poor prognosis.

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“…These findings have several important implications for cellular metabolism and human disease. The D-and L-isomers of HGA were previously detected in several organisms, including bacteria and humans, but the pathways leading to D-HGA and L-HGA synthesis were unknown (18,32). This report represents the first time that a route of HGA biosynthesis has been demonstrated.…”

Section: Discussionmentioning

confidence: 99%

A novel alpha-ketoglutarate reductase activity of the serA-encoded 3-phosphoglycerate dehydrogenase of Escherichia coli K-12 and its possible implications for human 2-hydroxyglutaric aciduria

1996

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Escherichia coli serA-encoded 3-phosphoglycerate (3PG) dehydrogenase catalyzes the first step of the major phosphorylated pathway of L-serine (Ser) biosynthesis. The SerA enzyme is evolutionarily related to the pdxB gene product, 4-phosphoerythronate dehydrogenase, which catalyzes the second step in one branch of pyridoxal 5-phosphate coenzyme biosynthesis. Both the Ser and pyridoxal 5-phosphate biosynthetic pathways use the serC(pdxF)-encoded transaminase in their next steps. In an analysis of these parallel pathways, we attempted to couple the transaminase and dehydrogenase reactions in the reverse direction. Unexpectedly, we found that the SerA enzyme catalyzes a previously undetected reduction of ␣-ketoglutarate (␣KG) to 2-hydroxyglutaric acid (HGA). Numerous criteria ruled out the possibility that this SerA ␣KG reductase activity was caused by contamination in the substrate or purified enzyme preparations. HGA was confirmed as the product of the

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Isolation and Characterization of a Mammalian Hydroxyacid-Oxoacid Transhydrogenase

Cited by 32 publications

References 13 publications

Disease-related Metabolites in Culture Medium of Fibroblasts from Patients with d-2-Hydroxyglutaric Aciduria, l-2-Hydroxyglutaric Aciduria, and Combined d/l-2-Hydroxyglutaric Aciduria

Disease-related Metabolites in Culture Medium of Fibroblasts from Patients with d-2-Hydroxyglutaric Aciduria, l-2-Hydroxyglutaric Aciduria, and Combined d/l-2-Hydroxyglutaric Aciduria

MYC-driven accumulation of 2-hydroxyglutarate is associated with breast cancer prognosis

A novel alpha-ketoglutarate reductase activity of the serA-encoded 3-phosphoglycerate dehydrogenase of Escherichia coli K-12 and its possible implications for human 2-hydroxyglutaric aciduria

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