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PMID: 19903557 Published · ppublish English Journal Article Research Support, N.I.H., Extramural

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

Methods in enzymology ·Vol. 464 ·2009-00-00 ·Pages 211-31

Ritchie TK, Grinkova YV, Bayburt TH, Denisov IG, Zolnerciks JK, Atkins WM, Sligar SG

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.

MeSH Terms
ATP Binding Cassette Transporter, Subfamily B, Member 1/chemistry Animals Bacteriorhodopsins/chemistry Crystallography, X-Ray Lipid Bilayers/chemistry Membrane Proteins/chemistry Mice Models, Biological Nanostructures/chemistry Phosphatidylcholines/chemistry Phospholipids/chemistry
Chemicals
ATP Binding Cassette Transporter, Subfamily B, Member 1 Lipid Bilayers Membrane Proteins Phosphatidylcholines Phospholipids Bacteriorhodopsins 1-palmitoyl-2-oleoylphosphatidylcholine
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Ritchie T K
Department of Medicinal Chemistry, University of Washington, Seattle, Washington, USA.
Grinkova Y V
Bayburt T H
Denisov I G
Zolnerciks J K
Atkins W M
Sligar S G
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Article Info
Journal
Methods in enzymology
Abbr.
Methods Enzymol
ISSN
1557-7988
DOI
10.1016/S0076-6879(09)64011-8
Published
2009-00-00
Pages
211-31
Language
English
Region
United States
NLM ID
0212271
PMCID
PMC4196316
Subset
IM
Grants
NIGMS NIH HHS · GM 32165 · United States
NIGMS NIH HHS · P01 GM032165 · United States
NIGMS NIH HHS · GM 31756 · United States
NIGMS NIH HHS · R01 GM031756 · United States
NIGMS NIH HHS · R37 GM031756 · United States
NIGMS NIH HHS · GM 33775 · United States
NIGMS NIH HHS · R01 GM033775 · United States
External Links
PubMed source DOI: 10.1016/S0076-6879(09)64011-8 PMC Full Text
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