InVivo EPR Characterization of Semi-Synthetic [FeFe] Hydrogenases
InVivo EPR Characterization of Semi-Synthetic [FeFe] Hydrogenases
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DOI：
10.1002/anie.201710740
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发表时间：
2018-03-01
影响因子：
16.6
通讯作者：
Berggren, Gustav
中科院分区：
文献类型：
--
作者：
Meszaros, Livia S.;Nemeth, Brigitta;Berggren, Gustav
EPR光谱揭示了两种不同的半合成氢化酶在体内的形成。[FeFe]氢化酶是催化分子氢和质子相互转化的金属酶。该反应由典型的铁硫簇和有机金属[2Fe]亚位组成的h簇催化。最近的研究表明，该酶可以用模拟[2Fe]亚位组成的合成辅因子进行重组，从而产生半合成的氢化酶。在这里，我们使用EPR光谱来监测这两种半合成酶在整个细胞中的形成。该研究首次提供了半合成氢化酶在体内的光谱表征，并在细胞内条件下观察了h簇的两种不同氧化状态。此外，这些发现强调了合成化学如何成为在体内条件下操纵和检查氢化酶的有力工具。
EPR spectroscopy reveals the formation of two different semi-synthetic hydrogenases invivo. [FeFe] hydrogenases are metalloenzymes that catalyze the interconversion of molecular hydrogen and protons. The reaction is catalyzed by the H-cluster, consisting of a canonical iron-sulfur cluster and an organometallic [2Fe] subsite. It was recently shown that the enzyme can be reconstituted with synthetic cofactors mimicking the composition of the [2Fe] subsite, resulting in semi-synthetic hydrogenases. Herein, we employ EPR spectroscopy to monitor the formation of two such semi-synthetic enzymes in whole cells. The study provides the first spectroscopic characterization of semi-synthetic hydrogenases invivo, and the observation of two different oxidized states of the H-cluster under intracellular conditions. Moreover, these findings underscore how synthetic chemistry can be a powerful tool for manipulation and examination of the hydrogenase enzyme under invivo conditions.