PDBsum entry 5oef

Oxidoreductase

PDB id: 5oef

Contents

Protein chains

574 a.a.

Ligands

9SQ ×2

SF4 ×8

FES ×2

Metals

_MG ×5

Waters ×716

PDB id:

5oef

Name:

Oxidoreductase

Title:

Active semisynthetic [fefe]-hydrogenase cpi with aza-diselenato- bridged [2fe] cofactor

Structure:

Iron hydrogenase 1. Chain: a, b. Synonym: cpi,fe-only hydrogenase,[fe] hydrogenase. Engineered: yes

Source:

Clostridium pasteurianum. Organism_taxid: 1501. Expressed in: escherichia coli bl21(de3). Expression_system_taxid: 469008. Expression_system_variant: discr

Resolution:

2.05Å R-factor: 0.234 R-free: 0.269

Authors:

L.Kertess,J.Esselborn,T.Happe,E.Hofmann

Key ref:

L.Kertess et al. (2017). Chalcogenide substitution in the [2Fe] cluster of [FeFe]-hydrogenases conserves high enzymatic activity. Dalton Trans, 46, 16947-16958. PubMed id: 29177350 DOI: 10.1039/c7dt03785f

Date:

07-Jul-17 Release date: 29-Nov-17

Protein chains

P29166  (PHF1_CLOPA) -  Iron hydrogenase 1 from Clostridium pasteurianum

Seq: 574 a.a.

Struc: 574 a.a.

Enzyme reactions

• Enzyme class: E.C.1.12.7.2 - ferredoxin hydrogenase.
• Reaction: H2 + 2 oxidized [2Fe-2S]-[ferredoxin] = 2 reduced [2Fe-2S]-[ferredoxin] + 2 H⁺
• Cofactor: Iron-sulfur; Ni(2+)

Molecule diagrams generated from .mol files obtained from the KEGG ftp site

Key reference

DOI no: 10.1039/c7dt03785f

Dalton Trans 46:16947-16958 (2017)

PubMed id: 29177350

Chalcogenide substitution in the [2Fe] cluster of [FeFe]-hydrogenases conserves high enzymatic activity.

L.Kertess, F.Wittkamp, C.Sommer, J.Esselborn, O.Rüdiger, E.J.Reijerse, E.Hofmann, W.Lubitz, M.Winkler, T.Happe, U.P.Apfel.

ABSTRACT

[FeFe]-Hydrogenases efficiently catalyze the uptake and evolution of H₂due to the presence of an inorganic [6Fe-6S]-cofactor (H-cluster). This cofactor is comprised of a [4Fe-4S] cluster coupled to a unique [2Fe] cluster where the catalytic turnover of H₂/H⁺takes place. We herein report on the synthesis of a selenium substituted [2Fe] cluster [Fe₂{μ(SeCH₂)₂NH}(CO)₄(CN)₂]^2-(ADSe) and its successful in vitro integration into the native protein scaffold of [FeFe]-hydrogenases HydA1 from Chlamydomonas reinhardtii and CpI from Clostridium pasteurianum yielding fully active enzymes (HydA1-ADSe and CpI-ADSe). FT-IR spectroscopy and X-ray structure analysis confirmed the presence of structurally intact ADSe at the active site. Electrochemical assays reveal that the selenium containing enzymes are more biased towards hydrogen production than their native counterparts. In contrast to previous chalcogenide exchange studies, the S to Se exchange herein is not based on a simple reconstitution approach using ionic cluster constituents but on the in vitro maturation with a pre-synthesized selenium-containing [2Fe] mimic. The combination of biological and chemical methods allowed for the creation of a novel [FeFe]-hydrogenase with a [2Fe2Se]-active site which confers individual catalytic features.