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

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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
          Methods Enzymol
          Methods in enzymology
          Elsevier BV
          1557-7988
          0076-6879
          2009
          : 464
          Affiliations
          [1 ] Department of Medicinal Chemistry, University of Washington, Seattle, Washington, USA.
          Article
          S0076-6879(09)64011-8 NIHMS634011
          10.1016/S0076-6879(09)64011-8
          4196316
          19903557
          753a11b4-59a7-4fb4-840b-51a21b3b775a
          History

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