Chapter 11 Reconstitution of Membrane Proteins in Phospholipid Bilayer Nanodiscs

T. K. Ritchie, Y. V. Grinkova, T. H. Bayburt, I. G. Denisov, J. K. Zolnerciks, W. M. Atkins, S. G. Sligar

Research output: Contribution to journalReview article

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.

Original languageEnglish (US)
Pages (from-to)211-231
Number of pages21
JournalMethods in enzymology
Volume464
Issue numberC
DOIs
StatePublished - Dec 14 2009

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology

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    Ritchie, T. K., Grinkova, Y. V., Bayburt, T. H., Denisov, I. G., Zolnerciks, J. K., Atkins, W. M., & Sligar, S. G. (2009). Chapter 11 Reconstitution of Membrane Proteins in Phospholipid Bilayer Nanodiscs. Methods in enzymology, 464(C), 211-231. https://doi.org/10.1016/S0076-6879(09)64011-8