Sequential and structural analysis of [NiFe]-hydrogenase-maturation proteins from Desulfovibrio vulgaris Miyazaki F
- 81 Downloads
The complete primary structure of the hyn-region in the genome of Desulfovibrio vulgaris Miyazaki F (DvMF), encoding the [NiFe]-hydrogenase and two maturation proteins has been identified. Besides the formerly reported genes for the large and small subunits, this region comprises genes encoding an endopeptidase (HynC) and a putative chaperone (HynD). The complete genomic region covers 4086 nucleotides including the previously published upstream located promoter region and the sequences of the structural genes. A phylogenetic tree for both maturation proteins shows strongest sequential relationship to the orthologous proteins of Desulfovibrio vulgaris Hildenborough (DvH). Secondary structure prediction for HynC (168 aa, corresponding to a molecular weight of 17.9 kDa) revealed a practically identical arrangement of α-helical and β-strand elements between the orthologous protein HybD from E. coli and allowed a three-dimensional modelling of HynC on the basis of the formerly published structure of HybD. The putative chaperone HynD consists of 83 aa (molecular weight of 9 kDa) and shows 76% homology to DvH HynD. Preliminary experiments demonstrate that the operon is expressed under the control of its own promoter in Escherichia coli, although no further processing could be observed, providing evidence that additional proteins have to be involved in the maturation process. Accession numbers: DQ072852, HynC protein ID AAY90127, HynD protein ID AAY90128.
KeywordsEndopeptidase [NiFe]-hydrogenase Operon structure Protein maturation process Three dimensional structure
Desulfovibrio vulgaris Hildenborough
Desulfovibrio vulgaris Miyazaki F
Unable to display preview. Download preview PDF.
The support of Ulrich Krauss, U. Düsseldorf/FZ Jülich, in the structure-modelling of HynC is greatly acknowledged. We wish to thank also Dr. Reiner Hedderich from the Max-Planck-Institute for Terrestrial Microbiology, Marburg, for valuable suggestions during the experimental work. This work was supported by the Max-Planck-Society.
- Brecht M, van Gastel M, Buhrke T, Friedrich B, Lubitz W (2003) Direct detection of a hydrogen ligand in the [NiFe] center of the regulatory H2-sensing hydrogenase from Ralstonia eutropha in its reduced state by HYSCORE and ENDOR spectroscopy. J Am Chem Soc 125:13075–13083PubMedCrossRefGoogle Scholar
- Heidelberg JF, Seshadri R, Haveman SA, Hemme CL, Paulsen IT, Kolonay JF, Eisen JA, Ward N, Methe B, Brinkac LM, Daugherty SC, Deboy RT, Dodson RJ, Durkin AS, Madupu R, Nelson WC, Sullivan SA, Fouts D, Haft DH, Selengut J, Peterson JD, Davidsen TM, Zafar N, Zhou LW, Radune D, Dimitrov G, Hance M, Tran K, Khouri H, Gill J, Utterback TR, Feldblyum TV, Wall JD, Voordouw G, Fraser CM (2004) The genome sequence of the anaerobic, sulfate-reducing bacterium Desulfovibrio vulgaris Hildenborough. Nat Biotechnol 22:554–559PubMedCrossRefGoogle Scholar
- Nicolet Y, De Lacey AL, Vernede X, Fernandez VM, Hatchikian EC, Fontecilla-Camps JC (2001) Crystallographic and FTIR spectroscopic evidence of changes in Fe coordination upon reduction of the active site of the Fe-only hydrogenase from Desulfovibrio desulfuricans. J Am Chem Soc 123:1596–1601PubMedCrossRefGoogle Scholar
- Vanderzwaan JW, Coremans JMCC, Bouwens ECM, Albracht SPJ (1990) Effect of O-17(2) and (Co)-C-13 on EPR-spectra of nickel in hydrogenase from Chromatium vinosum. Biochim Biophys Acta 1041:101–110Google Scholar