Home Immunology & Inflammation ROS activator Disodium (R)-2-Hydroxyglutarate

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Disodium (R)-2-Hydroxyglutarate ROS activator

Cat.No.S7873

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.
Disodium (R)-2-Hydroxyglutarate ROS activator Chemical Structure

Chemical Structure

Molecular Weight: 192.08

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Quality Control

Batch: Purity: 99.74%
99.74

Solubility

In vitro
Batch:

Water : 38 mg/mL

DMSO : Insoluble
(Moisture-contaminated DMSO may reduce solubility. Use fresh, anhydrous DMSO.)

Ethanol : Insoluble

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In vivo
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Chemical Information, Storage & Stability

Molecular Weight 192.08 Formula

C5H6Na2O5

 Storage (From the date of receipt)
CAS No. 103404-90-6 Download SDF Storage of Stock Solutions

Synonyms D-α-Hydroxyglutaric acid disodium Smiles C(CC(=O)[O-])C(C(=O)[O-])O.[Na+].[Na+]

Mechanism of Action

Targets/IC50/Ki
α-ketoglutarate-dependent dioxygenase
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. Besides, (R)-2-Hydroxyglutarate inhibits ATP synthase and mTOR signaling, and thus causes growth arrest and tumor cell killing.
Kinase Assay
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.
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