Novel type II and monomeric NAD+ specific isocitrate dehydrogenases: phylogenetic affinity, enzymatic characterization, and evolutionary implication.
Novel type II and monomeric NAD+ specific isocitrate dehydrogenases: phylogenetic affinity, enzymatic characterization, and evolutionary implication.
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新型 II 型和单体 NAD 特异性异柠檬酸脱氢酶：系统发育亲和力、酶学特征和进化意义。
DOI：
10.1038/srep09150
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发表时间：
2015-03-16
影响因子：
4.6
通讯作者：
Zhu G
中科院分区：
文献类型：
--
作者：
Wang P;Lv C;Zhu G
使用NAD+是异柠檬酸脱氢酶（IDH）的祖先特征，NADP+表型作为一个古老的适应事件在进化中出现。然而，在II型IDHs和单体IDHs中未发现NAD+特异性IDHs。本研究首次报道了来自lucimarinus Ostreococcus CCE9901 IDH （OlIDH）和Micromonas sp. RCC299 （MiIDH）的新型II型同型二聚体nadh - idhs，以及来自Campylobacter sp. FOBRC14 IDH （CaIDH）和弯曲弯曲杆菌（cidh）的新型单体nadh - idhs。采用尺寸排斥色谱法和MALDI-TOF/TOF质谱法测定同二聚体OlIDH和单体cidh。所有四种idh都被证明是NAD+特异性的，因为OlIDH、MiIDH、CaIDH和cidh对NAD+的偏好分别是NAD+的99倍、224倍、61倍和37倍。对推测的辅酶区分氨基酸（OlIDH中为Asp326/Met327， CaIDH中为Leu584/Asp595）进行了评价，发现两个突变体OlIDH R326H327和CaIDH H584R595的辅酶特异性从NAD+完全逆转为NADP+。进一步研究了OlIDH和cidh的详细生化特性，包括最佳反应pH和温度、热稳定性和金属离子效应。并对OlIDH、cidh和其他idh的进化联系进行了详细的描述和讨论。
NAD+ use is an ancestral trait of isocitrate dehydrogenase (IDH) and the NADP+ phenotype arose through evolution as an ancient adaptation event. However, no NAD+-specific IDHs have been found among type II IDHs and monomeric IDHs. In this study, novel type II homodimeric NAD-IDHs from Ostreococcus lucimarinus CCE9901 IDH (OlIDH) and Micromonas sp. RCC299 (MiIDH) and novel monomeric NAD-IDHs from Campylobacter sp. FOBRC14 IDH (CaIDH) and Campylobacter curvus (CcIDH) were reported for the first time. The homodimeric OlIDH and monomeric CaIDH were determined by size exclusion chromatography and MALDI-TOF/TOF mass spectrometry. All the four IDHs were demonstrated to be NAD+-specific, since OlIDH, MiIDH, CaIDH and CcIDH displayed 99-fold, 224-fold, 61-fold and 37-fold preferences for NAD+ over NADP+, respectively. The putative coenzyme discriminating amino acids (Asp326/Met327 in OlIDH, Leu584/Asp595 in CaIDH) were evaluated and the coenzyme specificities of the two mutants, OlIDH R326H327 and CaIDH H584R595, were completely reversed from NAD+ to NADP+. The detailed biochemical properties, including optimal reaction pH and temperature, thermostability and metal ion effects, of OlIDH and CaIDH were further investigated. The evolutionary connections among OlIDH, CaIDH and all the other forms of IDHs were described and discussed thoroughly.
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影响因子：
64.8
作者：
通讯作者：
--
影响因子：
64.8
作者：
Lu, Chao;Ward, Patrick S.;Kapoor, Gurpreet S.;Rohle, Dan;Turcan, Sevin;Abdel-Wahab, Omar;Edwards, Christopher R.;Khanin, Raya;Figueroa, Maria E.;Melnick, Ari;Wellen, Kathryn E.;O'Rourke, Donald M.;Berger, Shelley L.;Chan, Timothy A.;Levine, Ross L.;Mellinghoff, Ingo K.;Thompson, Craig B.
通讯作者：
Thompson, Craig B.
影响因子：
4.8
作者：
Hu, Gang;Lin, An-Ping;McAlister-Henn, Lee
通讯作者：
McAlister-Henn, Lee
影响因子：
4.8
作者：
Ceccarelli, C;Grodsky, NB;Bahnson, BJ
通讯作者：
Bahnson, BJ
影响因子：
3.9
作者：
Lu, Qian;Minard, Karyl I.;McAlister-Henn, Lee
通讯作者：
McAlister-Henn, Lee