Several intrinsically disordered proteins (IDPs) exhibit high affinity for lipid membranes. Among the different biophysical methods to probe protein-lipid interaction, neutron reflectometry (NR) can provide direct and structural detailed information on the location of the IDP with respect to the membrane. Supported lipid bilayers are commonly used as cell membrane models in such experiments. NR measurements can be collected on the supported lipid bilayer before and after the interaction with the IDP to characterize whether the protein molecules are mainly located on the membrane surface (interaction with the lipid headgroups), are penetrating into the hydrophobic region of the membrane (interaction with the lipid acyl chains), or are not interacting at all with the membrane. The lipid composition of the supported lipid bilayer can easily be tuned; hence the NR experiments can be designed to investigate selective IDP-lipid interactions.This chapter will describe the fundamental steps for performing an NR experiment and the subsequent data analysis aimed at characterizing IDP-lipid bilayer interactions. The specific case of an intrinsically disordered region (IDR) from the membrane protein Na+/H+ exchanger isoform 1 (NHE1) will be used as an example, but the same protocol can be easily adapted to other IDPs.
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