DOI > 10.5291/ILL-DATA.8-05-415

This proposal is publicly available since 08/02/2018

Title

Probing membrane protein structure using neutron reflectivity

Abstract

Membrane proteins represent about 30% of the proteomes of organisms and are dramatically under-represented in the structural database of the Protein Data Bank. This can be explained by their lack of stability and difficulty to crystallize. To answer this demand we propose to use a bacterial cell-free system to express hydrogenated and deuterated membrane proteins directly into a supported lipid bilayer in a neutron reflectivity (NR) solid-liquid cell. Cell-free systems are in vitro protein synthesis systems that use the extract (or lysate) from a prokaryotic or eukaryotic organism to provide the cellular machinery necessary for protein production. Here, we intend to use supported lipid bilayers to provide the lipidic environment into which the expressed membrane proteins will be directly inserted. To establish a proof of concept and demonstrate the potential of this new technique, we will investigate VDAC, a well characterized protein. Following these results, we will proceed to investigate the structure of Hepatitis C virus (HCV) proteins which have not, at this point, been crystallized.

Experimental Report

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Data Citation

The recommended format for citing this dataset in a research publication is in the following format:

WATKINS Erik; MARTIN Donald and SORANZO Thomas. (2013). Probing membrane protein structure using neutron reflectivity. Institut Laue-Langevin (ILL) doi:10.5291/ILL-DATA.8-05-415

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Metadata

Experiment Parameters

Sample Parameters

  • Formula

    • cell-free expression lysate
    • DMPC
  • Consistence

    solution