Background
L-α-Hydroxyglutaric Acid is an important metabolite in various domains of life. In mammals and plants, it is produced by lactate dehydrogenase (LDH) and malate dehydrogenase (MDH)-mediated 2-ketoglutarate (2-KG) reduction under hypoxic conditions[2]. L-α-Hydroxyglutaric Acid is an inhibitor of 2-KG dependent dioxygenases with specific pro-oncogenic capabilities[3,4].
In CD8-positive T-lymphocytes, L-α-Hydroxyglutaric Acid inhibited the production of effector cytokines and reduced cytotoxicity. In addition, when the dose of L-α-Hydroxyglutaric Acid was more than 300 μM, L-α-Hydroxyglutaric Acid inhibited cell expansion, increased cell apoptosis, decreased IFN-γ secretion, but increased IL-2 secretion[6].This compound was identified to aid the proliferation and antitumorigenic abilities of CD8+ T-lymphocytes, to contribute to relieving the cellular reductive stress[5].
When examined the effect of L-α-Hydroxyglutaric Acid, at concentrations varying from 1 to 5 mM, on CK activity in total homogenates from cerebellum, cerebral cortex, skeletal muscle and cardiac muscle. L-α-Hydroxyglutaric Acid did not alter tCK activities from cerebral cortex, skeletal muscle, and cardiac muscle, but significantly inhibited the enzyme activity from cerebellum at all concentrations tested. The inhibition of Mi-CK activity by L-α-Hydroxyglutaric Acid is non-competitive. The Km calculated was 2.52±0.74 mM. The Ki value was calculated by the method of which provides a simple way of determining the inhibition constant (Ki) for non-competitive inhibitors. The Ki value calculated was 11.13±3.71 mM for L-α-Hydroxyglutaric Acid [1].
Besides, in Zebrafish, L-α-Hydroxyglutaric Acid-induced apoptosis was caused by oxidation[7].
参考文献:
[1]: da Silva CG, Bueno AR, et,al. L-2-hydroxyglutaric acid inhibits mitochondrial creatine kinase activity from cerebellum of developing rats. Int J Dev Neurosci. 2003 Jun;21(4):217-24. doi: 10.1016/s0736-5748(03)00035-2. PMID: 12781789.
[2]: Oldham WM, Clish CB, et,al. Hypoxia-Mediated Increases in L-2-hydroxyglutarate Coordinate the Metabolic Response to Reductive Stress. Cell Metab. 2015 Aug 4;22(2):291-303. doi: 10.1016/j.cmet.2015.06.021. Epub 2015 Jul 23. PMID: 26212716; PMCID: PMC4526408.
[3]: Xu W, Yang H, et,al. Oncometabolite 2-hydroxyglutarate is a competitive inhibitor of α-ketoglutarate-dependent dioxygenases. Cancer Cell. 2011 Jan 18;19(1):17-30. doi: 10.1016/j.ccr.2010.12.014. PMID: 21251613; PMCID: PMC3229304.
[4]: Chowdhury R, Yeoh KK, et,al. The oncometabolite 2-hydroxyglutarate inhibits histone lysine demethylases. EMBO Rep. 2011 May;12(5):463-9. doi: 10.1038/embor.2011.43. Epub 2011 Apr 1. PMID: 21460794; PMCID: PMC3090014.
[5]: Tyrakis PA, Palazon A, et,al. S-2-hydroxyglutarate regulates CD8+ T-lymphocyte fate. Nature. 2016 Dec 8;540(7632):236-241. doi: 10.1038/nature20165. Epub 2016 Oct 26. PMID: 27798602; PMCID: PMC5149074.
[6]: Tyrakis PA, Palazon A, et,al. S-2-hydroxyglutarate regulates CD8+ T-lymphocyte fate. Nature. 2016 Dec 8;540(7632):236-241. doi: 10.1038/nature20165. Epub 2016 Oct 26. PMID: 27798602; PMCID: PMC5149074.
[7]:Parng C, Ton C, et,al. A zebrafish assay for identifying neuroprotectants in vivo. Neurotoxicol Teratol. 2006 Jul-Aug;28(4):509-16. doi: 10.1016/j.ntt.2006.04.003. Epub 2006 Jun 30. PMID: 16814516.
L-α-羟基戊二酸是生命各个领域的重要代谢产物。在哺乳动物和植物中，它是在低氧条件下通过乳酸脱氢酶 (LDH) 和苹果酸脱氢酶 (MDH) 介导的 2-酮戊二酸 (2-KG) 还原产生的[2]。 L-α-Hydroxyglutaric Acid 是一种 2-KG 依赖性双加氧酶抑制剂，具有特定的促癌能力[3,4]。
在 CD8 阳性 T 淋巴细胞中，L-α-羟基戊二酸可抑制效应细胞因子的产生并降低细胞毒性。此外，当L-α-羟基戊二酸的剂量大于300 μM时，L-α-羟基戊二酸抑制细胞增殖，增加细胞凋亡，减少IFN-γ分泌，但增加IL-2分泌[6 ].该化合物被鉴定为有助于 CD8+ T 淋巴细胞的增殖和抗肿瘤发生能力，有助于缓解细胞还原应激[5]。
当检查浓度从 1 到 5 mM 不等的 L-α-羟基戊二酸对来自小脑、大脑皮层、骨骼肌和心肌的总匀浆中 CK 活性的影响时。 L-α-Hydroxyglutaric Acid 不改变大脑皮层、骨骼肌和心肌的 tCK 活性，但在所有测试浓度下显着抑制小脑的酶活性。 L-α-Hydroxyglutaric Acid 对 Mi-CK 活性的抑制是非竞争性的。计算的 Km 为 2.52±0.74 mM。 Ki 值的计算方法提供了一种简单的方法来确定非竞争性抑制剂的抑制常数 (Ki)。 L-α-羟基戊二酸[1]的Ki值为11.13±3.71 mM。
此外，在斑马鱼中，L-α-羟基戊二酸诱导的细胞凋亡是由氧化引起的[7]。
Protocol
| Cell experiment [1]: |
|
Cell lines
|
CD8-positive T-lymphocytes
|
|
Preparation Method
|
Cells were activated with anti-CD3 and anti-CD28 antibodies for 48 h, then expanded for an additional 4 days in the presence of IL-2 and then treated with the indicated concentration of L-α-Hydroxyglutaric Acid for 16 h.
|
|
Reaction Conditions
|
300,500 μM L-α-Hydroxyglutaric Acid for 16 h
|
|
Applications
|
L-α-Hydroxyglutaric Acid inhibited the production of effector cytokines and reduced cytotoxicity. In addition, when the dose of L-α-Hydroxyglutaric Acid was more than 300 μM, L-α-Hydroxyglutaric Acid inhibited cell expansion, increased cell apoptosis, decreased IFN-γ secretion, but increased IL-2 secretion.
|
| Animal experiment [2]: |
|
Animal models
|
Wistar rats
|
|
Preparation Method
|
The homogenates of different mouse tissues were mixed with the corresponding buffer, and L-α-Hydroxyglutaric Acid at a final concentration of 0.25 to 5 mM was also added to the medium.
|
|
Dosage form
|
0.25 to 5 mM L-α-Hydroxyglutaric Acid for 15 min at 37 °C
|
|
Applications
|
When examined the effect of L-α-Hydroxyglutaric Acid, at concentrations varying from 1 to 5 mM, on CK activity in total homogenates from cerebellum, cerebral cortex, skeletal muscle and cardiac muscle. L-α-Hydroxyglutaric Acid did not alter tCK activities from cerebral cortex, skeletal muscle, and cardiac muscle, but significantly inhibited the enzyme activity from cerebellum at all concentrations tested.
|
|
参考文献:
[1]. Tyrakis PA, Palazon A, et,al. S-2-hydroxyglutarate regulates CD8+ T-lymphocyte fate. Nature. 2016 Dec 8;540(7632):236-241. doi: 10.1038/nature20165. Epub 2016 Oct 26. PMID: 27798602; PMCID: PMC5149074.
[2]. da Silva CG, Bueno AR, et,al. L-2-hydroxyglutaric acid inhibits mitochondrial creatine kinase activity from cerebellum of developing rats. Int J Dev Neurosci. 2003 Jun;21(4):217-24. doi: 10.1016/s0736-5748(03)00035-2. PMID: 12781789.
|
没有评价数据