• 主题
高级搜索  >
历史搜索 清空历史
首页> 美国卫生研究院文献>Chemical Science >Nuclear resonance vibrational spectroscopy reveals the FeS cluster composition and active site vibrational properties of an O2-tolerant NAD+-reducing NiFe hydrogenase
【2h】

Nuclear resonance vibrational spectroscopy reveals the FeS cluster composition and active site vibrational properties of an O2-tolerant NAD+-reducing NiFe hydrogenase

机译：核共振振动光谱揭示了耐O2的NAD +还原NiFe氢化酶的FeS簇组成和活性位点振动特性

代理获取
本网站仅为用户提供外文OA文献查询和代理获取服务，本网站没有原文。下单后我们将采用程序或人工为您竭诚获取高质量的原文，但由于OA文献来源多样且变更频繁，仍可能出现获取不到、文献不完整或与标题不符等情况，如果获取不到我们将提供退款服务。请知悉。
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

Hydrogenases are complex metalloenzymes that catalyze the reversible splitting of molecular hydrogen into protons and electrons essentially without overpotential. The NAD+-reducing soluble hydrogenase (SH) from Ralstonia eutropha is capable of H2 conversion even in the presence of usually toxic dioxygen. The molecular details of the underlying reactions are largely unknown, mainly because of limited knowledge of the structure and function of the various metal cofactors present in the enzyme. Here, all iron-containing cofactors of the SH were investigated by 57Fe specific nuclear resonance vibrational spectroscopy (NRVS). Our data provide experimental evidence for one [2Fe2S] center and four [4Fe4S] clusters, which is consistent with the amino acid sequence composition. Only the [2Fe2S] cluster and one of the four [4Fe4S] clusters were reduced upon incubation of the SH with NADH. This finding explains the discrepancy between the large number of FeS clusters and the small amount of FeS cluster-related signals as detected by electron paramagnetic resonance spectroscopic analysis of several NAD+-reducing hydrogenases. For the first time, Fe–CO and Fe–CN modes derived from the [NiFe] active site could be distinguished by NRVS through selective 13C labeling of the CO ligand. This strategy also revealed the molecular coordinates that dominate the individual Fe–CO modes. The present approach explores the complex vibrational signature of the Fe–S clusters and the hydrogenase active site, thereby showing that NRVS represents a powerful tool for the elucidation of complex biocatalysts containing multiple cofactors.
机译：氢酶是复杂的金属酶，可催化分子氢可逆地分裂成质子和电子，基本上没有超电势。来自富营养小球藻的NAD + 还原型可溶性氢化酶（SH）即使在通常有毒的双氧存在下也能够进行H 2转化。潜在反应的分子细节在很大程度上是未知的，主要是由于对酶中存在的各种金属辅因子的结构和功能的了解有限。在这里，通过 57 Fe特定核共振振动光谱法（NRVS）研究了SH的所有含铁辅因子。我们的数据提供了一个[2Fe2S]中心和四个[4Fe4S]簇的实验证据，这与氨基酸序列组成一致。 SH与NADH一起孵育后，只有[2Fe2S]簇和四个[4Fe4S]簇之一减少。这一发现解释了通过对几种NAD + 还原氢酶进行电子顺磁共振波谱分析所检测到的大量FeS簇与少量FeS簇相关信号之间的差异。 NRVS可以通过选择性地对CO配体进行 13 C标记来区分源自[NiFe]活性位点的Fe–CO和Fe–CN模式。该策略还揭示了主导单个Fe–CO模式的分子坐标。本方法探索了Fe–S团簇和氢化酶活性位点的复杂振动特征，从而表明NRVS代表了阐明含有多种辅助因子的复杂生物催化剂的强大工具。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
代理获取

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号