An EPR/HYSCORE, M?ssbauer, and resonance Raman study of the hydrogenase maturation enzyme HydF: a model for N-coordination to [4Fe-S] clusters
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- 作者：Gustav Berggren (1) (6)
Ricardo Garcia-Serres (2)
Xavier Brazzolotto (1)
Martin Clemancey (2)
Serge Gambarelli (3)
Mohamed Atta (1)
Jean-Marc Latour (2)
Heather L. Hernández (4)
Sowmya Subramanian (4)
Michael K. Johnson (4)
Marc Fontecave (1) (5) - 关键词：HydF ; Metalloenzyme ; Iron–sulfur cluster ; EPR ; M?ssbauer ; Hyperfine sublevel correlation
- 刊名：Journal of Biological Inorganic Chemistry
- 出版年：2014
- 出版时间：January 2014
- 年：2014
- 卷：19
- 期：1
- 页码：75-84
- 全文大小：434 KB
- 作者单位：Gustav Berggren (1) (6)
Ricardo Garcia-Serres (2)
Xavier Brazzolotto (1)
Martin Clemancey (2)
Serge Gambarelli (3)
Mohamed Atta (1)
Jean-Marc Latour (2)
Heather L. Hernández (4)
Sowmya Subramanian (4)
Michael K. Johnson (4)
Marc Fontecave (1) (5)
1. Laboratoire de Chimie et Biologie des Métaux, équipe??Biocatalyse?, Institut de Recherches en Technologies et Sciences pour le Vivant, iRTSV-LCBM/Biocat, UMR 5249 CEA/CNRS/UJF, CEA/Grenoble, 17, rue des Martyrs, Grenoble, France
6. Department of Biochemistry and Biophysics, Stockholm University, Svante Arrhenius v?g 16, 106 91, Stockholm, Sweden
2. Laboratoire de Chimie et Biologie des Métaux, équipe?“Physicochimie des Métaux en Biologie- Institut de Recherches en Technologies et Sciences pour le Vivant, iRTSV-LCBM/pmb, UMR 5249 CEA/CNRS/UJF, CEA/Grenoble, Grenoble, France
3. Laboratoire?“Résonance Magnétique- Université Joseph Fourier, Grenoble 1/CEA/Institut Nanoscience et Cryogénie/SCIB, UMR-E3, Grenoble, France
4. Department of Chemistry and Center for Metalloenzyme Studies, University of Georgia, Athens, GA, 30602, USA
5. Collège de France, 11 place Marcellin-Berthelot, Paris, France - ISSN：1432-1327
文摘
The biosynthesis of the organometallic H cluster of [Fe–Fe]?hydrogenase requires three accessory proteins, two of which (HydE and HydG) belong to the radical S-adenosylmethionine enzyme superfamily. The third, HydF, is an Fe–S protein with GTPase activity. The [4Fe-S] cluster of HydF is bound to the polypeptide chain through only the three, conserved, cysteine residues present in the binding sequence motif CysXHisX(46-53)HisCysXXCys. However, the involvement of the two highly conserved histidines as a fourth ligand for the cluster coordination is controversial. In this study, we set out to characterize further the [4Fe-S] cluster of HydF using M?ssbauer, EPR, hyperfine sublevel correlation (HYSCORE), and resonance Raman spectroscopy in order to investigate the influence of nitrogen ligands on the spectroscopic properties of [4Fe-S]2+/+ clusters. Our results show that M?ssbauer, resonance Raman, and EPR spectroscopy are not able to readily discriminate between the imidazole-coordinated [4Fe-S] cluster and the non-imidazole-bound [4Fe-S] cluster with an exchangeable fourth ligand that is present in wild-type HydF. HYSCORE spectroscopy, on the other hand, detects the presence of an imidazole/histidine ligand on the cluster on the basis of the appearance of a specific spectral pattern in the strongly coupled region, with a coupling constant of approximately 6?MHz. We also discovered that a His-tagged version of HydF, with a hexahistidine tag at the N-terminus, has a [4Fe-S] cluster coordinated by one histidine from the tag. This observation strongly indicates that care has to be taken in the analysis of data obtained on tagged forms of metalloproteins.