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Complementing structural information of modular proteins with small angle neutron scattering and contrast variation

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An Erratum to this article was published on 26 March 2008

Abstract

Many macromolecules in the cell function by forming multi-component assemblies. We have applied the technique of small angle neutron scattering to study a nucleic acid–protein complex and a multi-protein complex. The results illustrate the versatility and applicability of the method to study macromolecular assemblies. The neutron scattering experiments, complementing X-ray solution scattering data, reveal that the conserved catalytic domain of RNase E, an essential ribonuclease in Escherichia coli (E. coli), undergoes a marked conformational change upon binding a 5′monophosphate–RNA substrate analogue. This provides the first evidence in support of an allosteric mechanism that brings about RNA substrate cleavage. Neutron contrast variation of the multi-protein TIM10 complex, a mitochondrial chaperone assembly comprising the subunits Tim9 and Tim10, has been used to determine a low-resolution shape reconstruction of the complex, highlighting the integral subunit organization. It shows characteristic features involving protrusions that could be assigned to the six subunits forming the complex.

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Abbreviations

SANS:

Small angle neutron scattering

SAXS:

Small angle X-ray scattering

RNase E :

Ribonuclease E

TIM10:

Translocase of the intermembrane space

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Acknowledgments

The work on RNase E was supported by the Wellcome Trust. We thank Martyn Symmons and Martin Moncrieffe for stimulating discussions and invaluable advice. The work on TIM10 was supported by funds from IMBB-FORTH. The ILL and STFC Daresbury Laboratory are acknowledged for beamtime. We are also very grateful to Peter Timmins at the ILL and Michael Haertlein at the ILL-EMBL-PSB Deuteration Laboratory in Grenoble for their excellent support.

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Author notes

  1. A. J. Callaghan

    Present address: Biophysics Laboratories, Institute of Biomedical and Biomolecular Science, University of Portsmouth, King Henry Building, King Henry 1st Street, Portsmouth, PO1 2DY, UK

Authors and Affiliations

  1. Molecular Biophysics Group, STFC Daresbury Laboratory, Daresbury Science and Innovation Campus, Warrington, Cheshire, WA4 AD, UK

    J. G. Grossmann

  2. Department of Biochemistry, University of Cambridge, 80 Tennis Court Road, Cambridge, CB2 1GA, UK

    A. J. Callaghan, M. J. Marcaida & B. F. Luisi

  3. Institute of Molecular Biology and Biotechnology, Foundation for Research and Technology Hellas, PO Box 1385, Heraklion, 71110, Crete, Greece

    F. H. Alcock & K. Tokatlidis

Authors
  1. J. G. Grossmann
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  2. A. J. Callaghan
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  3. M. J. Marcaida
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  4. B. F. Luisi
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  5. F. H. Alcock
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  6. K. Tokatlidis
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Corresponding author

Correspondence to J. G. Grossmann.

Additional information

Advanced neutron scattering and complementary techniques to study biological systems. Contributions from the meetings, “Neutrons in Biology”, STFC Rutherford Appleton Laboratory, Didcot, UK, 11–13 July and “Proteins At Work 2007”, Perugia, Italy, 28–30 May 2007.

An erratum to this article can be found at http://dx.doi.org/10.1007/s00249-008-0302-3

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Grossmann, J.G., Callaghan, A.J., Marcaida, M.J. et al. Complementing structural information of modular proteins with small angle neutron scattering and contrast variation. Eur Biophys J 37, 603–611 (2008). https://doi.org/10.1007/s00249-008-0278-z

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