8CJY

[FeFe]-hydrogenase CpI from Clostridium pasteurianum, variant S357T

8CJY の概要

• エントリーDOI: 10.2210/pdb8cjy/pdb
• 関連するPDBエントリー: 4XDC
• 分子名称: Iron hydrogenase 1, dicarbonyl[bis(cyanide-kappaC)]-mu-(iminodimethanethiolatato-1kappaS:2kappaS)-mu-(oxomethylidene)diiron(2+), IRON/SULFUR CLUSTER, ... (8 entities in total)
• 機能のキーワード: [fefe]-hydrogenase, cpi from clostridium pasteurianum, variant s357t, oxidoreductase
• 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: 134548.75

構造登録者

Brocks, C.,Duan, J.,Hofmann, E.,Happe, T. (登録日: 2023-02-13, 公開日: 2023-10-25, 最終更新日: 2024-02-21)

主引用文献

Brocks, C.,Das, C.K.,Duan, J.,Yadav, S.,Apfel, U.P.,Ghosh, S.,Hofmann, E.,Winkler, M.,Engelbrecht, V.,Schafer, L.V.,Happe, T.
A Dynamic Water Channel Affects O 2 Stability in [FeFe]-Hydrogenases.
Chemsuschem, 17:e202301365-e202301365, 2024

PubMed Abstract: [FeFe]-hydrogenases are capable of reducing protons at a high rate. However, molecular oxygen (O ) induces the degradation of their catalytic cofactor, the H-cluster, which consists of a cubane [4Fe4S] subcluster (4Fe ) and a unique diiron moiety (2Fe ). Previous attempts to prevent O -induced damage have focused on enhancing the protein's sieving effect for O by blocking the hydrophobic gas channels that connect the protein surface and the 2Fe . In this study, we aimed to block an O diffusion pathway and shield 4Fe instead. Molecular dynamics (MD) simulations identified a novel water channel (W ) surrounding the H-cluster. As this hydrophilic path may be accessible for O molecules we applied site-directed mutagenesis targeting amino acids along W in proximity to 4Fe to block O diffusion. Protein film electrochemistry experiments demonstrate increased O stabilities for variants G302S and S357T, and MD simulations based on high-resolution crystal structures confirmed an enhanced local sieving effect for O in the environment of the 4Fe in both cases. The results strongly suggest that, in wild type proteins, O diffuses from the 4Fe to the 2Fe . These results reveal new strategies for improving the O stability of [FeFe]-hydrogenases by focusing on the O diffusion network near the active site.

PubMed: 37830175
DOI: 10.1002/cssc.202301365

実験手法

X-RAY DIFFRACTION (1.6 Å)