Kinetic and inhibition studies on human Jumonji-C (JmjC) domain-containing protein 5.
Kinetic and inhibition studies on human Jumonji-C (JmjC) domain-containing protein 5.
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DOI：
10.1039/d2cb00249c
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发表时间：
2023-06-07
影响因子：
4.1
通讯作者：
Schofield, Christopher J.
中科院分区：
文献类型：
--
作者：
Tumber, Anthony;Salah, Eidarus;Brewitz, Lennart;Corner, Thomas P.;Schofield, Christopher J.
Jumonji-C(JmjC)结构域包含蛋白5(JMJD5)是一种依赖于人的2-氧戊二酸(2OG)和铁(II)的加氧酶，它催化翻译后精氨酸残基的C3羟基化，并通过未知的机制与昼夜节律和癌症生物学联系在一起。我们报告了基于固相萃取-质谱仪(SPE-MS)的JMJD5分析方法，该方法能够进行动力学和高通量的抑制研究。动力学研究表明，一些人工合成的2OG衍生物，特别是带有环碳骨架的2OG衍生物(即(1R)-3-(carboxycarbonyl)cyclopentane-1-carboxylic酸)，是JMJD5和抑制低氧诱导转录因子α(FIH)的有效替代底物，但不是Jumonji-C(JMJC)组蛋白Nε-甲基赖氨酸去甲基酶KDM4E的替代底物，这显然反映了JMJD5和FIH结构上的更接近的相似性。通过考察已报道的2OG加氧酶抑制剂对JMJD5的催化作用，验证了JMJD5抑制实验的有效性；结果表明，广谱的2OG加氧酶抑制剂也是有效的JMJD5抑制剂(如N-草甘氨酸、吡啶-2，4-二羧酸、ebselen)，而大多数临床使用的2OG加氧酶抑制剂(如roxadustat)不抑制JMJD5。SPE-MS分析将有助于开发高效和选择性的JMJD5抑制剂，用于研究JMJD5在细胞研究中的生化功能。报道了一种基于稳健的质谱学分析方法，用于监测2-氧-戊二酸依赖的加氧酶Jumonji-C结构域蛋白5(JMJD5)的催化作用。这些分析方法已应用于动力学和抑制研究。
Jumonji-C (JmjC) domain-containing protein 5 (JMJD5) is a human 2-oxoglutarate (2OG) and Fe(ii)-dependent oxygenase which catalyses the post-translational C3 hydroxylation of arginyl-residues and which is linked to the circadian rhythm and to cancer biology through as yet unidentified mechanisms. We report robust solid phase extraction coupled to mass spectrometry (SPE-MS)-based JMJD5 assays which enable kinetic and high-throughput inhibition studies. The kinetic studies reveal that some synthetic 2OG derivatives, notably including a 2OG derivative with a cyclic carbon backbone (i.e. (1R)-3-(carboxycarbonyl)cyclopentane-1-carboxylic acid), are efficient alternative cosubstrates of JMJD5 and of factor inhibiting hypoxia-inducible transcription factor HIF-α (FIH), but not of the Jumonji-C (JmjC) histone Nε-methyl lysine demethylase KDM4E, apparently reflecting the closer structural similarity of JMJD5 and FIH. The JMJD5 inhibition assays were validated by investigating the effect of reported 2OG oxygenase inhibitors on JMJD5 catalysis; the results reveal that broad-spectrum 2OG oxygenase inhibitors are also efficient JMJD5 inhibitors (e.g. N-oxalylglycine, pyridine-2,4-dicarboxylic acid, ebselen) whereas most 2OG oxygenase inhibitors that are in clinical use (e.g. roxadustat) do not inhibit JMJD5. The SPE-MS assays will help enable the development of efficient and selective JMJD5 inhibitors for investigating the biochemical functions of JMJD5 in cellular studies. Robust mass spectrometry-based assays are reported which monitor catalysis of the 2-oxoglutarate-dependent oxygenase Jumonji-C domain-containing protein 5 (JMJD5). The assays have been applied to both kinetic and inhibition studies.
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DOI：
10.1002/anie.201706788
发表时间：
2017-12-04
期刊：
Angewandte Chemie (International ed. in English)
影响因子：
--
作者：
Gerken PA;Wolstenhulme JR;Tumber A;Hatch SB;Zhang Y;Müller S;Chandler SA;Mair B;Li F;Nijman SMB;Konietzny R;Szommer T;Yapp C;Fedorov O;Benesch JLP;Vedadi M;Kessler BM;Kawamura A;Brennan PE;Smith MD
通讯作者：
Smith MD
影响因子：
3.4
作者：
Beck H;Jeske M;Thede K;Stoll F;Flamme I;Akbaba M;Ergüden JK;Karig G;Keldenich J;Oehme F;Militzer HC;Hartung IV;Thuss U
通讯作者：
Thuss U
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：
4.5
作者：
Amendola PG;Zaghet N;Ramalho JJ;Vilstrup Johansen J;Boxem M;Salcini AE
通讯作者：
Salcini AE
影响因子：
5.3
作者：
Del Rizzo, Paul A.;Krishnan, Swathi;Trievel, Raymond C.
通讯作者：
Trievel, Raymond C.