The structure of the Ni–Fe site in the isolated HoxC subunit of the hydrogen‐sensing hydrogenase from Ralstonia eutropha
- 18 July 2005
- journal article
- Published by Wiley in FEBS Letters
- Vol. 579 (20) , 4287-4291
- https://doi.org/10.1016/j.febslet.2005.06.063
Abstract
The regulatory Ni–Fe hydrogenase (RH) from Ralstonia eutropha which forms a [HoxBC]2 complex functions as a hydrogen sensor under aerobic conditions. We have studied a novel Strep-tag isolate of the RH large subunit, HoxCST, which lacks the Fe–S clusters of HoxB, allowing for structure determination of the catalytic site by X-ray absorption spectroscopy both at the Ni and, for the first time, also at the Fe K-edge. This technique, together with Fourier-transform infrared spectroscopy, revealed a Ni–Fe site with [O1(CysS)2NiII(μ-SCys)2FeII(CN)2(CO)] structure in about 50% of HoxCST and a [(CysS)2FeII(CN)2(CO)] site lacking Ni in the remainder protein. Possibly both sites may be intermediates in the maturation process of the RHKeywords
This publication has 22 references indexed in Scilit:
- A model system for [NiFe] hydrogenase maturation studies: Purification of an active site‐containing hydrogenase large subunit without small subunitFEBS Letters, 2005
- Reduction of Unusual Iron-Sulfur Clusters in the H2-sensing Regulatory Ni-Fe Hydrogenase from Ralstonia eutropha H16Journal of Biological Chemistry, 2005
- Structural and Oxidation-State Changes at Its Nonstandard Ni−Fe Site during Activation of the NAD-Reducing Hydrogenase from Ralstonia eutropha Detected by X-ray Absorption, EPR, and FTIR SpectroscopyJournal of the American Chemical Society, 2004
- Direct Detection of a Hydrogen Ligand in the [NiFe] Center of the Regulatory H2-Sensing Hydrogenase fromRalstoniaeutrophain Its Reduced State by HYSCORE and ENDOR SpectroscopyJournal of the American Chemical Society, 2003
- Hydrogen-Induced Structural Changes at the Nickel Site of the Regulatory [NiFe] Hydrogenase from Ralstonia eutropha Detected by X-ray Absorption SpectroscopyBiochemistry, 2003
- X-ray absorption spectroscopy to analyze nuclear geometry and electronic structure of biological metal centers?potential and questions examined with special focus on the tetra-nuclear manganese complex of oxygenic photosynthesisAnalytical and Bioanalytical Chemistry, 2003
- The H2 Sensor of Ralstonia eutrophaPublished by Elsevier ,2001
- Crystal structure of the nickel–iron hydrogenase from Desulfovibrio gigasNature, 1995
- Absolute energy calibration of X-ray radiation from synchrotron sourcesJournal of Applied Crystallography, 1985
- A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye bindingAnalytical Biochemistry, 1976