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Solution NMR Spectroscopy for the Determination of Structures of Membrane Proteins in a Lipid Environment

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Lipid-Protein Interactions

Part of the book series: Methods in Molecular Biology ((MIMB,volume 974))

Abstract

Several recent advancements have transformed solution NMR spectroscopy into a competitive, elegant, and eminently viable technique for determining the solution structures of membrane proteins at the level of atomic resolution. Once a good level of cell-based or cell-free expression and purification of a suitably sized membrane protein has been achieved, then NMR offers a combination of several versatile strategies, for example, choice of appropriate deuterated or non-deuterated detergents, temperature, and ionic strength; isotope labelling with 2H, 13C, 15N, with or without protonation of Ile (δ1), Leu, and Val methyl protons; combinatorial labelling of specific amino acids; transverse relaxation-optimized NMR spectroscopy-based, Nonuniform sampling-based, and other NMR experiments; measurement of residual dipolar couplings using stretched polyacrylamide gels or DNA nanotubes; and spin-labelling and paramagnetic relaxation enhancements. Strategic combinations of these advancements together with availability of highly sensitive cryogenically cooled probes equipped high-field NMR spectrometers (up to 1 GHz 1H frequency) have allowed the perseverant investigator to successfully overcome several of the conventional pitfalls associated with the NMR technique and membrane proteins, viz., low sensitivity, poor sample stability, spectral crowding, and a limited number of NOEs and other constraints for structure calculations. This has resulted in an unprecedented growth in the number of successfully determined NMR structures of large and complex membrane proteins, and this technique now holds great promise for the structure determination of an ever larger body of membrane proteins.

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Author information

Authors and Affiliations

  1. Molecular and Structural Biology, CSIR-Central Drug Research Institute, Lucknow, UP, India

    Ashish Arora

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  1. Ashish Arora
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Correspondence to Ashish Arora .

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Editors and Affiliations

  1. Institut für Biologie, Abteilung Biophysik, Universität Kassel, Heinrich-Plett-Straße 40, Kassel, 34132, Germany

    Jörg H. Kleinschmidt

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Arora, A. (2013). Solution NMR Spectroscopy for the Determination of Structures of Membrane Proteins in a Lipid Environment. In: Kleinschmidt, J. (eds) Lipid-Protein Interactions. Methods in Molecular Biology, vol 974. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-275-9_17

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  • DOI: https://doi.org/10.1007/978-1-62703-275-9_17

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-62703-274-2

  • Online ISBN: 978-1-62703-275-9

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