Disodium (R)-2-Hydroxyglutarate

Catalog No.S7873 Batch:S787304

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Technical Data

Formula

C5H6Na2O5

Molecular Weight 192.08 CAS No. 103404-90-6
Solubility (25°C)* In vitro Water 38 mg/mL (197.83 mM)
DMSO Insoluble
Ethanol Insoluble
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

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Biological Activity

Description Disodium (R)-2-Hydroxyglutarate (D-α-Hydroxyglutaric acid disodium) is a competitive inhibitor of α-ketoglutarate-dependent dioxygenases with Ki of 10.87 ± 1.85 mM.
Targets
α-ketoglutarate-dependent dioxygenase [1]
In vitro In U-87MG cells, (R)-2-Hydroxyglutarate acts as weak antagonists of α-KG to inhibit α-KG-dependent histone demethylases and increases dimethylation on both H3K9 and H3K79. [1] Besides, (R)-2-Hydroxyglutarate inhibits ATP synthase and mTOR signaling, and thus causes growth arrest and tumor cell killing. [2]

Protocol (from reference)

Kinase Assay:

[1]

  • Enzymatic Assays

    To assay human JHDM1A/KDM2A demethylase activity toward H3K36me2, His tagged JHDM1A is first obtained by transforming pET28a-JHDM1A into Escherichia coli BL21 and protein expression is induced by addition of 1 mM IPTG at 30° C when cell density reaches 0.5 OD600 units. Cells are lysed by sonication and Ni-NTA agarose is used to purify His-JHDM1A fusion proteins. Histone demethylase assay is carried out by incubating 2 μg oligonucleosomes, 4 μg purified His-JHDM1A, and/or 10–50 mM L- or D-2-HG in histone demethylation buffer [50 mM HEPES (pH 8.0), 625 μM Fe(NH4)2(SO4)2, 0.1–0.5 mM α-KG, 2 mM ascorbate] at 37° C for 2–3 hr and the reactions are stopped by the addition of SDS loading buffer and subsequently analyzed by western blotting using anti-H3K36me2 antibody. To measure CeKDM7A demethylase activity toward H3K9me2 and H3K27me2, two synthetic dimethylated peptides H3K9me2 [ARTKQTARK (me2)STGGKA] and H3K27me2 [QLATKAARK (me2)SAPAS] are used as substrates. Demethylase assays are carried out in the presence of 10 μg enzyme, 1 μg peptide in 20 μl buffer 20 mM Tris-HCl (pH 7.5), 150 mM NaCl, 50 μM Fe(NH4)2(SO4)22, 100 μM α-KG, 2 mM Vc, 10 mM PMSF for 3 hr. The demethylation reaction mixture is desalted by passing through a C18 ZipTip. To examine the inhibitory effect of 2-HG, various concentrations of 2-HG are incubated with KDM7A briefly before adding other reaction mixtures. The samples are analyzed by a MALDI-TOF/TOF mass spectrometer.

Cell Assay:

[2]

  • Cell lines

    U87/IDH1(R132H), Octyl α-KG- or octyl 2-HG-treated U87, and HCT 116 IDH1(R132H/+) cells

  • Concentrations

    ~800 μM

  • Incubation Time

    4 days

  • Method

    Cells are seeded in 12-well plates and, after overnight incubation, treated with the indicated concentrations of each compound. After harvesting, cells are stained with acridine orange (AO) and 4′,6-diamidino-2-phenylindole (DAPI). Cell number and viability are measured based on AO and DAPI fluorescence as measured by NC3000 following the manufacturer’s instructions.

Selleck's Disodium (R)-2-Hydroxyglutarate has been cited by 7 publications

Itaconate ameliorates autoimmunity by modulating T cell imbalance via metabolic and epigenetic reprogramming [ Nat Commun, 2023, 14(1):984] PubMed: 36849508
Induction of functional neutrophils from mouse fibroblasts by thymidine through enhancement of Tet3 activity [ Cell Mol Immunol, 2022, 10.1038/s41423-022-00842-9] PubMed: 35301470
D2HGDH-mediated D2HG catabolism enhances the anti-tumor activities of CAR-T cells in an immunosuppressive microenvironment [ Mol Ther, 2022, S1525-0016(22)00007-7] PubMed: 35007759
NAD+ Metabolism Maintains Inducible PD-L1 Expression to Drive Tumor Immune Evasion [ Cell Metab, 2021, 33(1):110-127.e5] PubMed: 33171124
Wild-Type IDH1 and Mutant IDH1 Opposingly Regulate Podoplanin Expression in Glioma [ Transl Oncol, 2020, 13(4):100758] PubMed: 32208352
Isocitrate dehydrogenase1 mutation reduces the pericyte coverage of microvessels in astrocytic tumours [ J Neurooncol, 2019, 143(2):187-196] PubMed: 31004262
Cytokine correlation analysis based on drug perturbation [Wallner FK, et al. Cytokine, 2016, 90:73-79] PubMed: 27816795

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

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