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Fundamental and biotechnological applications of neutron scattering measurements for macromolecular dynamics

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Abstract

To explore macromolecular dynamics on the picosecond timescale, we used neutron spectroscopy. First, molecular dynamics were analyzed for the hyperthermophile malate dehydrogenase from Methanococcus jannaschii and a mesophilic homologue, the lactate dehydrogenase from Oryctolagus cunniculus muscle. Hyperthermophiles have elaborate molecular mechanisms of adaptation to extremely high temperature. Using a novel elastic neutron scattering approach that provides independent measurements of the global flexibility and of the structural resilience (rigidity), we have demonstrated that macromolecular dynamics represents one of these molecular mechanisms of thermoadaptation. The flexibilities were found to be similar for both enzymes at their optimal activity temperature and the resilience is higher for the hyperthermophilic protein. Secondly, macromolecular motions were examined in a native and immobilized dihydrofolate reductase (DHFR) from Escherichia coli. The immobilized mesophilic enzyme has increased stability and decreased activity, so that its properties are changed to resemble those of the thermophilic enzyme. Are these changes reflected in dynamical behavior? For this study, we performed quasielastic neutron scattering measurements to probe the protein motions. The residence time is 7.95 ps for the native DHFR and 20.36 ps for the immobilized DHFR. The average height of the potential barrier to local motions is therefore increased in the immobilized DHFR, with a difference in activation energy equal to 0.54 kcal/mol, which is, using the theoretical rate equation, of the same order than expected from calculation.

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Acknowledgments

M.T. was supported by the Région Rhône-Alpes, France, the Marsden Fund of the Royal Society of New Zealand, New Zealand, and by the Istituto Nazionale Fisica della Materia, Italy. We acknowledge supports from the EU under the DLAB contracts HPRI-CT-2001-50035 and RII3-CT-2003-505925, and the CNRS GEOMEX programme. We thank the CRG-IN13 and the Institut Laue Langevin for support and for providing the neutron facilities used for this work.

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Authors and Affiliations

  1. CNR-INFM, c/o Institut Laue-Langevin, 6 rue Jules Horowitz, BP 156, 38042, Grenoble Cedex 9, France

    Moeava Tehei

  2. Department of Biological Sciences, University of Waikato, Private Bag 3105, Hamilton, New Zealand

    Roy Daniel

  3. Institut Laue-Langevin, 6 rue Jules Horowitz, BP 156, 38042, Grenoble Cedex 9, France

    Giuseppe Zaccai

  4. Institut de Biologie Structurale, UMR 5075 CEA-CNRS-UJF, 41 rue Jules Horowitz, 38027, Grenoble, France

    Giuseppe Zaccai

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  1. Moeava Tehei
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Correspondence to Moeava Tehei.

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Tehei, M., Daniel, R. & Zaccai, G. Fundamental and biotechnological applications of neutron scattering measurements for macromolecular dynamics. Eur Biophys J 35, 551–558 (2006). https://doi.org/10.1007/s00249-006-0082-6

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  • DOI: https://doi.org/10.1007/s00249-006-0082-6

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