Identification and characterization of a noncanonical menaquinone-linked formate dehydrogenase.
Identification and characterization of a noncanonical menaquinone-linked formate dehydrogenase.
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DOI：
10.1016/j.jbc.2021.101384
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发表时间：
2022-03
期刊：
影响因子：
--
通讯作者：
Walburger A
中科院分区：
文献类型：
--
作者：
Arias-Cartín R;Uzel A;Seduk F;Gerbaud G;Pierrel F;Broc M;Lebrun R;Guigliarelli B;Magalon A;Grimaldi S;Walburger A
甲酸脱氢酶（FDHs）的钼/钨-双pyranopterin鸟嘌呤二核苷酸家族在从碳固定到能量收集的多种代谢途径中发挥作用，因为它们与各种氧化还原伙伴发生反应。事实上，这种代谢可塑性来自于与催化枢纽相互作用的伙伴亚基所使用的不同结构、辅因子含量和底物。在这里，我们揭示了枯草芽孢杆菌中的两个非规范FDHs，它们被组织成具有独特特征的两个亚基复合物，ForCE1和ForCE2。我们发现甲酸氧化还原酶催化亚基与一个前所未有的伙伴亚基，甲酸氧化还原酶必需亚基相互作用，并且其活性位点内的氨基酸序列偏离了通常与FDH活性相关的共识残基，因为组氨酸残基自然地被谷氨酰胺取代。甲酸氧化还原酶必需亚基介导了甲基萘醌作为电子受体的利用，这两种酶的甲酸甲萘醌氧化还原酶活性、它们与甲基萘醌的共化以及纯化后的酶通过多频电子顺磁共振（EPR）实验对蛋白质结合的中性甲基萘醌自由基的独特检测表明。此外，两种FDHs的EPR表征显示存在几个具有明显弛豫特性的[Fe-S]簇和弱各向异性Mo(V) EPR特征，这与该酶家族的钼/双pyranopterin鸟嘌呤二核苷酸辅因子的特征一致。总之，这项工作扩大了我们对FDH家族的认识，通过确定一个非规范的FDH，它在结构、钼辅助因子周围的氨基酸保护和反应性方面有所不同。
The molybdenum/tungsten—bis-pyranopterin guanine dinucleotide family of formate dehydrogenases (FDHs) plays roles in several metabolic pathways ranging from carbon fixation to energy harvesting because of their reaction with a wide variety of redox partners. Indeed, this metabolic plasticity results from the diverse structures, cofactor content, and substrates used by partner subunits interacting with the catalytic hub. Here, we unveiled two noncanonical FDHs in Bacillus subtilis, which are organized into two-subunit complexes with unique features, ForCE1 and ForCE2. We show that the formate oxidoreductase catalytic subunit interacts with an unprecedented partner subunit, formate oxidoreductase essential subunit, and that its amino acid sequence within the active site deviates from the consensus residues typically associated with FDH activity, as a histidine residue is naturally substituted with a glutamine. The formate oxidoreductase essential subunit mediates the utilization of menaquinone as an electron acceptor as shown by the formate:menadione oxidoreductase activity of both enzymes, their copurification with menaquinone, and the distinctive detection of a protein-bound neutral menasemiquinone radical by multifrequency electron paramagnetic resonance (EPR) experiments on the purified enzymes. Moreover, EPR characterization of both FDHs reveals the presence of several [Fe-S] clusters with distinct relaxation properties and a weakly anisotropic Mo(V) EPR signature, consistent with the characteristic molybdenum/bis-pyranopterin guanine dinucleotide cofactor of this enzyme family. Altogether, this work enlarges our knowledge of the FDH family by identifying a noncanonical FDH, which differs in terms of architecture, amino acid conservation around the molybdenum cofactor, and reactivity.
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DOI：
10.1038/nsb994
发表时间：
2003-11-01
期刊：
NATURE STRUCTURAL BIOLOGY
影响因子：
--
作者：
Arnoux, P;Sabaty, M;Pignol, D
通讯作者：
Pignol, D
影响因子：
16.6
作者：
Arnoux, Pascal;Ruppelt, Christian;Walburger, Anne
通讯作者：
Walburger, Anne
影响因子：
56.9
作者：
Boyington, JC;Gladyshev, VN;Sun, PD
通讯作者：
Sun, PD
影响因子：
2.9
作者：
BARBER, MJ;MAY, HD;FERRY, JG
通讯作者：
FERRY, JG
影响因子：
4.1
作者：
GANGESWARAN, R;LOWE, DJ;EADY, RR
通讯作者：
EADY, RR