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      Chapter 11 - Reconstitution of membrane proteins in phospholipid bilayer nanodiscs.

      Methods in enzymology

      Animals, Bacteriorhodopsins, chemistry, Crystallography, X-Ray, Lipid Bilayers, Membrane Proteins, Mice, Models, Biological, Nanostructures, P-Glycoprotein, Phosphatidylcholines, Phospholipids

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          Abstract

          Self-assembled phospholipid bilayer Nanodiscs have become an important and versatile tool among model membrane systems to functionally reconstitute membrane proteins. Nanodiscs consist of lipid domains encased within an engineered derivative of apolipoprotein A-1 scaffold proteins, which can be tailored to yield homogeneous preparations of disks with different diameters, and with epitope tags for exploitation in various purification strategies. A critical aspect of the self-assembly of target membranes into Nanodiscs lies in the optimization of the lipid:protein ratio. Here we describe strategies for performing this optimization and provide examples for reconstituting bacteriorhodopsin as a trimer, rhodopsin, and functionally active P-glycoprotein. Together, these demonstrate the versatility of Nanodisc technology for preparing monodisperse samples of membrane proteins of wide-ranging structure.

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          Author and article information

          Journal
          19903557
          10.1016/S0076-6879(09)64011-8
          4196316

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