Structure of human RNA N⁶-methyladenine demethylase ALKBH5 provides insights into its mechanisms of nucleic acid recognition and demethylation.
Structure of human RNA N⁶-methyladenine demethylase ALKBH5 provides insights into its mechanisms of nucleic acid recognition and demethylation.
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人RNAN⁶-甲基杜氏丁基脱甲基酶AlkBH5的结构提供了有关其核酸识别和脱甲基化机制的见解。
DOI：
10.1093/nar/gku085
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发表时间：
2014-04
影响因子：
14.9
通讯作者：
McDonough MA
中科院分区：
文献类型：
--
作者：
Aik W;Scotti JS;Choi H;Gong L;Demetriades M;Schofield CJ;McDonough MA
ALKBH 5是一种2-酮戊二酸（2 OG）和亚铁依赖性核酸加氧酶（NAOX），催化RNA中N6-甲基腺嘌呤的去甲基化。ALKBH 5在缺氧情况下上调，并在精子发生中发挥作用。我们描述了人ALKBH 5（残基66-292）的晶体结构，分辨率为2.0 nm。ALKBH 566 -292具有双链β-螺旋核心折叠，如在其他2 OG和铁依赖性加氧酶家族成员中观察到的。活性位点金属通过HXD. H基序（包含残基His 204、Asp 206和His 266）和三个水分子八面体配位。ALKBH 5与其他NAOX共享核苷酸识别盖和保守的活性位点残基。ALKBH 5中的一个大环（βIV-V）占据了与脂肪量和肥胖相关蛋白的L1环相似的区域，该区域被提议赋予单链RNA选择性。出乎意料的是，观察到小分子抑制剂IOX 3共价连接到位于活性位点外部的Cys 200的侧链。基于其他NAOX-核酸复合物将底物建模为活性位点揭示了对识别和去甲基化机制重要的保守残基。结构的见解将有助于开发NAOX选择性抑制剂，用作功能探针和治疗益处。
ALKBH5 is a 2-oxoglutarate (2OG) and ferrous iron-dependent nucleic acid oxygenase (NAOX) that catalyzes the demethylation of N6-methyladenine in RNA. ALKBH5 is upregulated under hypoxia and plays a role in spermatogenesis. We describe a crystal structure of human ALKBH5 (residues 66–292) to 2.0 Å resolution. ALKBH566–292 has a double-stranded β-helix core fold as observed in other 2OG and iron-dependent oxygenase family members. The active site metal is octahedrally coordinated by an HXD…H motif (comprising residues His204, Asp206 and His266) and three water molecules. ALKBH5 shares a nucleotide recognition lid and conserved active site residues with other NAOXs. A large loop (βIV–V) in ALKBH5 occupies a similar region as the L1 loop of the fat mass and obesity-associated protein that is proposed to confer single-stranded RNA selectivity. Unexpectedly, a small molecule inhibitor, IOX3, was observed covalently attached to the side chain of Cys200 located outside of the active site. Modelling substrate into the active site based on other NAOX–nucleic acid complexes reveals conserved residues important for recognition and demethylation mechanisms. The structural insights will aid in the development of inhibitors selective for NAOXs, for use as functional probes and for therapeutic benefit.
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影响因子：
8.4
作者：
Hopkinson RJ;Tumber A;Yapp C;Chowdhury R;Aik W;Che KH;Li XS;Kristensen JBL;King ONF;Chan MC;Yeoh KK;Choi H;Walport LJ;Thinnes CC;Bush JT;Lejeune C;Rydzik AM;Rose NR;Bagg EA;McDonough MA;Krojer T;Yue WW;Ng SS;Olsen L;Brennan PE;Oppermann U;Muller-Knapp S;Klose RJ;Ratcliffe PJ;Schofield CJ;Kawamura A
通讯作者：
Kawamura A
影响因子：
64.8
作者：
Han, Zhifu;Niu, Tianhui;Chai, Jijie
通讯作者：
Chai, Jijie
影响因子：
56.9
作者：
Choudhary, Chunaram;Kumar, Chanchal;Mann, Matthias
通讯作者：
Mann, Matthias
影响因子：
64.5
作者：
Chen, Zhongzhou;Zang, Jianye;Zhang, Gongyi
通讯作者：
Zhang, Gongyi
DOI：
10.1107/s0907444909052925
发表时间：
2010-02
期刊：
Acta crystallographica. Section D, Biological crystallography
影响因子：
--
作者：
Adams PD;Afonine PV;Bunkóczi G;Chen VB;Davis IW;Echols N;Headd JJ;Hung LW;Kapral GJ;Grosse-Kunstleve RW;McCoy AJ;Moriarty NW;Oeffner R;Read RJ;Richardson DC;Richardson JS;Terwilliger TC;Zwart PH
通讯作者：
Zwart PH