Structural differences between the ready and unready oxidized states of [NiFe] hydrogenases

  • Anne Volbeda
  • Lydie Martin
  • Christine Cavazza
  • Michaël Matho
  • Bart W. Faber
  • Winfried Roseboom
  • Simon P. J. Albracht
  • Elsa Garcin
  • Marc Rousset
  • Juan C. Fontecilla-Camps
Original Article

DOI: 10.1007/s00775-005-0632-x

Cite this article as:
Volbeda, A., Martin, L., Cavazza, C. et al. J Biol Inorg Chem (2005) 10: 239. doi:10.1007/s00775-005-0632-x

Abstract

[NiFe] hydrogenases catalyze the reversible heterolytic cleavage of molecular hydrogen. Several oxidized, inactive states of these enzymes are known that are distinguishable by their very different activation properties. So far, the structural basis for this difference has not been understood because of lack of relevant crystallographic data. Here, we present the crystal structure of the ready Ni-B state of Desulfovibrio fructosovorans [NiFe] hydrogenase and show it to have a putative μ-hydroxo Ni–Fe bridging ligand at the active site. On the other hand, a new, improved refinement procedure of the X-ray diffraction data obtained for putative unready Ni-A/Ni-SU states resulted in a more elongated electron density for the bridging ligand, suggesting that it is a diatomic species. The slow activation of the Ni-A state, compared with the rapid activation of the Ni-B state, is therefore proposed to result from the different chemical nature of the ligands in the two oxidized species. Our results along with very recent electrochemical studies suggest that the diatomic ligand could be hydro–peroxide.

Keywords

X-ray crystallography [NiFe] hydrogenase Oxidative inactivation Activation 

Abbreviations

DFT

Density functional theory

ENDOR

Electron–nuclear double resonance

EPR

Electron paramagnetic resonance

ESRF

European Synchrotron Radiation Facility

EXAFS

Extended X-ray absorption fine structure

FT

Fourier transform

PEG

Poly(ethylene glycol)

TLS

Translation–libration–screw

Copyright information

© SBIC 2005

Authors and Affiliations

  • Anne Volbeda
    • 1
  • Lydie Martin
    • 1
  • Christine Cavazza
    • 1
  • Michaël Matho
    • 1
  • Bart W. Faber
    • 2
  • Winfried Roseboom
    • 2
  • Simon P. J. Albracht
    • 2
  • Elsa Garcin
    • 1
  • Marc Rousset
    • 3
  • Juan C. Fontecilla-Camps
    • 1
  1. 1.Laboratoire de Cristallographie et de Cristallogenèse des ProtèinesInstitut de Biologie Structurale J.P. Ebel (CEA-CNRS-UJF)Grenoble Cédex 1France
  2. 2.Biochemistry, Swammerdam Institute for Life SciencesUniversity of AmsterdamAmsterdamThe Netherlands
  3. 3.Unité de Bioénergétique et Ingénierie des ProtéinesInstitut de Biologie Structurale et Microbiologie, CNRSMarseille Cédex 20France

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