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Reduced hybrid cluster proteins (HCP) from Desulfovibrio desulfuricans ATCC 27774 and Desulfovibrio vulgaris (Hildenborough): X-ray structures at high resolution using synchrotron radiation

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Abstract

The hybrid cluster proteins from the sulfate reducing bacteria Desulfovibrio desulfuricans ATCC 27774 (Dd) and Desulfovibrio vulgaris strain Hildenborough (Dv) have been isolated and crystallized anaerobically. In each case, the protein has been reduced with dithionite and the crystal structure of the reduced form elucidated using X-ray synchrotron radiation techniques at 1.25 Å and 1.55 Å resolution for Dd and Dv, respectively. Although the overall structures of the proteins are unchanged upon reduction, there are significant changes at the hybrid cluster centres. These include significant movements in the position of the iron atom linked to the persulfide moiety in the oxidized as-isolated proteins and the sulfur atom of the persulfide itself. The nature of these changes is described and the implications with respect to the function of hybrid cluster proteins are discussed.

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Fig. 1. a
Fig. 2. a
Fig. 3a, b.
Fig. 4.
Fig. 5.

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Abbreviations

Dd :

Desulfovibrio desulfuricans ATCC 27774

Dv :

Desulfovibrio vulgaris strain Hildenborough

DTN:

dithionite anion, S2O42−

GOL:

1,2,3-trihydroxypropane (glycerol)

HCP:

hybrid cluster protein, formerly known as prismane

MES:

2-(N-morpholino)ethanesulfonic acid

PEG:

poly(ethylene glycol)

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Acknowledgements

We thank the European Synchrotron Radiation Facility, Grenoble, France, and the staff of the Macromolecular Crystallography Group for the provision of synchrotron radiation facilities, without which the high-resolution studies would not have been possible. The ESRF also provided a PhD studentship for S.M. and kindly provided partial support for D.G.A. D.G.A. also acknowledges a grant from Fundação para a Ciência e Tecnologia (FCT), SFRH (BD/6480/2001). C.V.R. acknowledges a grant from Praxis XXI (BD/19879/99). P.F.L. would like to acknowledge the assistance and cooperation of the Department of Crystallography, Birkbeck College, University of London, UK. This work was partially funded by FCT, SAPIENS (POCTI/BME/38859/2001). The authors would also like to thank to Dora Alves for her valuable help on some aspects of Dv protein purification. Figures 1 to 4 were produced with the help of the programs Molscript 28, Bobscript 29and Raster 3D 30

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Authors and Affiliations

  1. Instituto de Tecnologia Química e Biológica, Universidade Nova de Lisboa, Av. República, Apartado 127, 2781-901 , Oeiras, Portugal

    David Aragão, Sofia Macedo, Célia V. Romão, Carlos Frazão, Lígia M. Saraiva, António V. Xavier, Miguel Teixeira, Maria A. Carrondo & Peter Lindley

  2. European Synchrotron Radiation Facility, BP 220, 38043 Cedex, Grenoble , France

    David Aragão, Sofia Macedo, Edward P. Mitchell & Peter Lindley

  3. Department of Biochemistry, University of Georgia, Athens, GA , 30602, USA

    Ming Y. Liu & Jean LeGall

  4. Department of Biochemistry, Wageningen University, Dreijenlaan 3, 6703 HA , Wageningen, The Netherlands

    Walter M. A. M. van Dongen

  5. Kluyver Department of Biotechnology, Delft University of Technology, Julianalaan 67, 2628 BC , Delft, The Netherlands

    Wilfred R. Hagen

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  1. David Aragão
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  2. Sofia Macedo
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  14. Peter Lindley
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Correspondence to Peter Lindley.

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Aragão, D., Macedo, S., Mitchell, E.P. et al. Reduced hybrid cluster proteins (HCP) from Desulfovibrio desulfuricans ATCC 27774 and Desulfovibrio vulgaris (Hildenborough): X-ray structures at high resolution using synchrotron radiation. J Biol Inorg Chem 8, 540–548 (2003). https://doi.org/10.1007/s00775-003-0443-x

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