[FeFe]-Hydrogenase with Chalcogenide Substitutions at the H-Cluster Maintains Full H2 Evolution Activity
[FeFe]-Hydrogenase with Chalcogenide Substitutions at the H-Cluster Maintains Full H2 Evolution Activity
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DOI：
10.1002/anie.201511896
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发表时间：
2016-07-11
影响因子：
16.6
通讯作者：
Happe, Thomas
中科院分区：
文献类型：
--
作者：
Noth, Jens;Esselborn, Julian;Happe, Thomas
来自莱茵衣藻的[FeFe]-氢化酶HYDA 1特别适合于生物化学和生物物理表征，因为活性位点中的H-簇是存在的唯一无机辅因子。在此，我们提出了将H-簇完全化学掺入HYDA 1-脱辅基蛋白支架中，此外，还成功地用硒取代了天然[4Fe（H）]簇中的硫。测定重构的未成熟HYDA 1-[4Fe 4Se]（H）蛋白的晶体结构，并在体外成熟时产生催化完整的人工H簇变体。通过动力学分析、FTIR光谱和X-射线结构分析，验证了重新设计的酶的完全析氢活性以及类似天然的组成和行为。这些发现表明，即使是具有特殊复杂性的生物无机活性位点也可以表现出令人惊讶的组成可塑性。
The [FeFe]-hydrogenase HYDA1 from Chlamydomonas reinhardtii is particularly amenable to biochemical and biophysical characterization because the H-cluster in the active site is the only inorganic cofactor present. Herein, we present the complete chemical incorporation of the H-cluster into the HYDA1-apoprotein scaffold and, furthermore, the successful replacement of sulfur in the native [4Fe(H)] cluster with selenium. The crystal structure of the reconstituted pre-mature HYDA1-[4Fe4Se](H) protein was determined, and a catalytically intact artificial H-cluster variant was generated upon in vitro maturation. Full hydrogen evolution activity as well as native-like composition and behavior of the redesigned enzyme were verified through kinetic assays, FTIR spectroscopy, and X-ray structure analysis. These findings reveal that even a bioinorganic active site with exceptional complexity can exhibit a surprising level of compositional plasticity.