Abstract
Neutron scattering has significant benefits for examining the structure of protein–lipid complexes. Cold (slow) neutrons are nondamaging and predominantly interact with the atomic nucleus, meaning that neutron beams can penetrate deeply into samples, which allows for flexibility in the design of samples studied. Most importantly, there is a strong difference in neutron scattering length (i.e., scattering power) between protium (\( {}_1{}^1H \), 99.98% natural abundance) and deuterium (\( {}_1{}^2H \) or D, 0.015%). Through the mixing of H2O and D2O in the samples and in some cases the deuterium labeling of the biomolecules, components within a complex can be hidden or enhanced in the scattering signal. This enables both the overall structure and the relative distribution of components within a complex to be resolved. Lipid–protein complexes are most commonly studied using neutron reflectometry (NR) and small angle neutron scattering (SANS). In this review the methodologies to produce and examine a variety of model biological membrane systems using SANS and NR are detailed. These systems include supported lipid bilayers derived from vesicle dispersions or Langmuir–Blodgett deposition, tethered bilayer systems, membrane protein–lipid complexes and polymer wrapped lipid nanodiscs. The three key stages of any SANS/NR study on model membrane systems—sample preparation, data collection, and analysis—are described together with some background on the techniques themselves.
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21 April 2020
This chapter was inadvertently published with the expansion of the term “MNB” printed incorrectly as “N-Methyl-N-nitrosobenzamide” under section 2.5. Instead, it should have been “Methyl 4- nitrobenzenesulfonate.” This correction has been updated in the chapter.
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Acknowledgments
L.A.C. would like to thank Max Skoda, Nicolò Paracini, Martynas Gavutis, Robert Dalgliesh, Sophie Ayscough, and John Webster for helpful discussions and advice; Filip Ciesielski for providing membrane structure diagrams; and Arwel Hughes for fitting software and associated error estimation functions.
Certain commercial materials, equipment, and instruments are identified in this work to describe the experimental procedure as completely as possible. In no case does such an identification imply a recommendation or endorsement by NIST, nor does it imply that the materials, equipment, or instrument identified are necessarily the best available for the purpose.
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Clifton, L.A., Hall, S.C.L., Mahmoudi, N., Knowles, T.J., Heinrich, F., Lakey, J.H. (2019). Structural Investigations of Protein–Lipid Complexes Using Neutron Scattering. In: Kleinschmidt, J. (eds) Lipid-Protein Interactions. Methods in Molecular Biology, vol 2003. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9512-7_11
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