5LA3

[FeFe]-hydrogenase CpI from Clostridium pasteurianum, variant E279A

5LA3 の概要

• エントリーDOI: 10.2210/pdb5la3/pdb
• 分子名称: Iron hydrogenase 1, dicarbonyl[bis(cyanide-kappaC)]-mu-(iminodimethanethiolatato-1kappaS:2kappaS)-mu-(oxomethylidene)diiron(2+), IRON/SULFUR CLUSTER, ... (6 entities in total)
• 機能のキーワード: hydrogenase, h-cluster, oxidoreductase, semisynthetic enzyme
• 由来する生物種: Clostridium pasteurianum
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 133767.63

構造登録者

Duan, J.,Esselborn, J.,Hofmann, E.,Winkler, M.,Happe, T. (登録日: 2016-06-13, 公開日: 2017-06-21, 最終更新日: 2024-01-10)

主引用文献

Winkler, M.,Senger, M.,Duan, J.,Esselborn, J.,Wittkamp, F.,Hofmann, E.,Apfel, U.P.,Stripp, S.T.,Happe, T.
Accumulating the hydride state in the catalytic cycle of [FeFe]-hydrogenases.
Nat Commun, 8:16115-16115, 2017

PubMed Abstract: H turnover at the [FeFe]-hydrogenase cofactor (H-cluster) is assumed to follow a reversible heterolytic mechanism, first yielding a proton and a hydrido-species which again is double-oxidized to release another proton. Three of the four presumed catalytic intermediates (H, H/H and H) were characterized, using various spectroscopic techniques. However, in catalytically active enzyme, the state containing the hydrido-species, which is eponymous for the proposed heterolytic mechanism, has yet only been speculated about. We use different strategies to trap and spectroscopically characterize this transient hydride state (H) for three wild-type [FeFe]-hydrogenases. Applying a novel set-up for real-time attenuated total-reflection Fourier-transform infrared spectroscopy, we monitor compositional changes in the state-specific infrared signatures of [FeFe]-hydrogenases, varying buffer pH and gas composition. We selectively enrich the equilibrium concentration of H, applying Le Chatelier's principle by simultaneously increasing substrate and product concentrations (H/H). Site-directed manipulation, targeting either the proton-transfer pathway or the adt ligand, significantly enhances H accumulation independent of pH.

PubMed: 28722011
DOI: 10.1038/ncomms16115

実験手法

X-RAY DIFFRACTION (2.29 Å)