Natural and synthetic 2-oxoglutarate derivatives are substrates for oncogenic variants of human isocitrate dehydrogenase 1 and 2.
Natural and synthetic 2-oxoglutarate derivatives are substrates for oncogenic variants of human isocitrate dehydrogenase 1 and 2.
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DOI：
10.1016/j.jbc.2023.102873
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发表时间：
2023-02
影响因子：
4.8
通讯作者：
Schofield, Christopher J.
中科院分区：
文献类型：
--
作者：
Liu, Xiao;Reinbold, Raphael;Liu, Shuang;Herold, Ryan A.;Rabe, Patrick;Duclos, Stephanie;Yadav, Rahul B.;Abboud, Martine I.;Thieffine, Sandrine;Armstrong, Fraser A.;Brewitz, Lennart;Schofield, Christopher J.
异柠檬酸脱氢酶(IDH)1和2的变异体(IDH1/2)通过催化NADPH依赖的2-氧戊二酸(2OG)还原为(2R)-羟基戊二酸来改变癌细胞的代谢。然而，目前还不清楚2OG的衍生物如何影响癌细胞的代谢。在这里，我们使用合成的C3和C4烷基化的2OG衍生物来研究最常见的癌症相关IDH1变体(R132H IDH1)、两个癌症相关IDH2变体(R172K IDH2、R140Q IDH2)和WT IDH1/2的底物选择性。在存在和不存在2OG的情况下，我们使用吸光度、核磁共振和电化学分析来监测WT IDH1/2和IDH1/2变体催化的2OG衍生物的周转。我们的结果表明，2OG衍生物可以作为所研究的IDH1/2变体的底物，但不能作为WT IDH1/2的底物，并且有可能作为20G竞争抑制物。动力学参数表明，一些2OG衍生物，包括天然产物3-甲基-2OG，是与2OG相同甚至更有效的IDH1/2变体底物。此外，核磁共振和质谱学研究证实，在3-甲基、3-丁基和3-苄基取代的2OG衍生物中，IDH1/2变体催化产生醇；带有IDH1变体(R132C/S280F IDH1)的3-丁基-2OG的晶体结构显示了活性中心结合。综合结果突出了(I)IDH1/2变体催化细胞内2OG以外的2-氧酸的还原，(Ii)2-氧酸天然产物(包括常见食品中的一些)对IDH1/2变体活性的调节，(Iii)通过活性结合而不是已建立的变构抑制模式来抑制IDH1/2变体，以及(Iv)IDH1/2变体作为生物催化剂的可能性。
Variants of isocitrate dehydrogenase (IDH) 1 and 2 (IDH1/2) alter metabolism in cancer cells by catalyzing the NADPH-dependent reduction of 2-oxoglutarate (2OG) to (2R)-hydroxyglutarate. However, it is unclear how derivatives of 2OG can affect cancer cell metabolism. Here, we used synthetic C3- and C4-alkylated 2OG derivatives to investigate the substrate selectivities of the most common cancer-associated IDH1 variant (R132H IDH1), of two cancer-associated IDH2 variants (R172K IDH2, R140Q IDH2), and of WT IDH1/2. Absorbance-based, NMR, and electrochemical assays were employed to monitor WT IDH1/2 and IDH1/2 variant-catalyzed 2OG derivative turnover in the presence and absence of 2OG. Our results reveal that 2OG derivatives can serve as substrates of the investigated IDH1/2 variants, but not of WT IDH1/2, and have the potential to act as 2OG-competitive inhibitors. Kinetic parameters reveal that some 2OG derivatives, including the natural product 3-methyl-2OG, are equally or even more efficient IDH1/2 variant substrates than 2OG. Furthermore, NMR and mass spectrometry studies confirmed IDH1/2 variant-catalyzed production of alcohols in the cases of the 3-methyl–, 3-butyl–, and 3-benzyl–substituted 2OG derivatives; a crystal structure of 3-butyl-2OG with an IDH1 variant (R132C/S280F IDH1) reveals active site binding. The combined results highlight the potential for (i) IDH1/2 variant-catalyzed reduction of 2-oxoacids other than 2OG in cells, (ii) modulation of IDH1/2 variant activity by 2-oxoacid natural products, including some present in common foods, (iii) inhibition of IDH1/2 variants via active site binding rather than the established allosteric mode of inhibition, and (iv) possible use of IDH1/2 variants as biocatalysts.
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DOI：
10.1021/acs.jpclett.1c01517
发表时间：
2021-07-08
期刊：
The journal of physical chemistry letters
影响因子：
--
作者：
Herold RA;Reinbold R;Megarity CF;Abboud MI;Schofield CJ;Armstrong FA
通讯作者：
Armstrong FA
影响因子：
5.2
作者：
Idle JR;Seipel K;Bacher U;Pabst T;Beyoğlu D
通讯作者：
Beyoğlu D
2-氧气谷物衍生物的合成及其作为人天冬氨酸/天冬酰胺-β-羟化酶的蛋白质和抑制剂的评估。
影响因子：
8.4
作者：
Brewitz L;Nakashima Y;Schofield CJ
通讯作者：
Schofield CJ
影响因子：
64.8
作者：
通讯作者：
--
影响因子：
3.8
作者：
Al-Qahtani, Khalid;Jabeen, Bushra;McCullagh, James S. O.
通讯作者：
McCullagh, James S. O.