Journal article
Characterization of a cyanobacterial-like uptake [NiFe] hydrogenase: EPR and FTIR spectroscopic studies of the enzyme from Acidithiobacillus ferrooxidans
Max-Volmer-Institut für Biophysikalische Chemie und Biochemie, Technische Universität Berlin, 10623, Berlin, Germany.1
Electron paramagnetic resonance (EPR) and Fourier transform IR studies on the soluble hydrogenase from Acidithiobacillus ferrooxidans are presented. In addition, detailed sequence analyses of the two subunits of the enzyme have been performed. They show that the enzyme belongs to a group of uptake [NiFe] hydrogenases typical for Cyanobacteria.
The sequences have also a close relationship to those of the H(2)-sensor proteins, but clearly differ from those of standard [NiFe] hydrogenases. It is concluded that the structure of the catalytic centre is similar, but not identical, to that of known [NiFe] hydrogenases. The active site in the majority of oxidized enzyme molecules, 97% in cells and more than 50% in the purified enzyme, is EPR-silent.
Upon contact with H(2) these sites remain EPR-silent and show only a limited IR response. Oxidized enzyme molecules with an EPR-detectable active site show a Ni(r)*-like EPR signal which is light-sensitive at cryogenic temperatures. This is a novelty in the field of [NiFe] hydrogenases. Reaction with H(2) converts these active sites to the well-known Ni(a)-C* state.
Illumination below 160 K transforms this state into the Ni(a)-L* state. The reversal, in the dark at 200 K, proceeds via an intermediate Ni EPR signal only observed with the H(2)-sensor protein from Ralstonia eutropha. The EPR-silent active sites in as-isolated and H(2)-treated enzyme are also light-sensitive as observed by IR spectra at cryogenic temperatures.
The possible origin of the light sensitivity is discussed. This study represents the first spectral characterization of an enzyme of the group of cyanobacterial uptake hydrogenases.
Language: | English |
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Publisher: | Springer-Verlag |
Year: | 2007 |
Pages: | 212-233 |
ISSN: | 14321327 and 09498257 |
Types: | Journal article |
DOI: | 10.1007/s00775-006-0185-7 |
Acidithiobacillus Acidithiobacillus ferrooxidans Amino Acid Sequence Biochemistry, general Cyanobacteria Electron Spin Resonance Spectroscopy Electron paramagnetic resonance and Fourier transform IR Hydrogen Hydrogenase Iron Iron-Sulfur Proteins Kinetics Life Sciences Light Light sensitivity Microbiology Nickel Oxidation-Reduction Spectroscopy, Fourier Transform Infrared Uptake [NiFe] hydrogenase nickel-iron hydrogenase