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Membrane Protein Structure Determination pp 261–282Cite as

Determination of Membrane Protein Structures Using Solution and Solid-State NMR

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 654))

Abstract

NMR is an essential tool to characterize the structure, dynamics, and interactions of biomolecules at an atomic level. Its application to membrane protein (MP) structure determination is challenging and currently an active and rapidly developing field. Main difficulties are the low sensitivity of the technique, the size limitation, and the intrinsic motional properties of the system under investigation. Solution and solid-state NMR (ssNMR) have common and own specific requirements. Solution NMR requires a careful choice of the detergent, elaborated stable isotope labelling schemes to overcome signal overlaps and to collect distance restraints. Excessive spectra crowding hampered large MP structure determination by ssNMR, and so far only high resolution structure of small or fragments of MP have been determined. However, ssNMR provides the unique opportunity to obtain atomic level information of MP in phospholipid bilayers such as orientation of the protein in the membrane. Specific and careful sample preparations are required in combination with uniformly and partially labelled protein for ssNMR spectra assignment. Distance restraints measurements benefit from methodologies currently developed for small soluble proteins in micro-crystalline state.

Recent advances in the field increased the releasing rate of high resolution MP structures, providing unprecedented structural and dynamics information making NMR a powerful tool for structural and functional membrane protein studies.

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  1. CEA, iBiTecs, URA 2096, SB2SM, Gif-sur-Yvette, France

    Pierre Montaville & Nadège Jamin

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  1. Pierre Montaville
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  2. Nadège Jamin
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Correspondence to Nadège Jamin .

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  1. , Université Paris Diderot - Paris 7, INSERM U773/CRB3, rue Henri Huchard 16, Paris, 75018, France

    Jean-Jacques Lacapère

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Montaville, P., Jamin, N. (2010). Determination of Membrane Protein Structures Using Solution and Solid-State NMR. In: Lacapère, JJ. (eds) Membrane Protein Structure Determination. Methods in Molecular Biology, vol 654. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-762-4_14

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  • DOI: https://doi.org/10.1007/978-1-60761-762-4_14

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  • Publisher Name: Humana Press, Totowa, NJ

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