Document Detail


Optimized phospholipid bilayer nanodiscs facilitate high-resolution structure determination of membrane proteins.
MedLine Citation:
PMID:  23294159     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Structural studies of membrane proteins are still hampered by difficulties of finding appropriate membrane-mimicking media that maintain protein structure and function. Phospholipid nanodiscs seem promising to overcome the intrinsic problems of detergent-containing environments. While nanodiscs can offer a near-native environment, the large particle size complicates their routine use in the structural analysis of membrane proteins by solution NMR. Here, we introduce nanodiscs assembled from shorter ApoA-I protein variants that are of markedly smaller diameter and show that the resulting discs provide critical improvements for the structure determination of membrane proteins by NMR. Using the bacterial outer-membrane protein OmpX as an example, we demonstrate that the combination of small nanodisc size, high deuteration levels of protein and lipids, and the use of advanced non-uniform NMR sampling methods enable the NMR resonance assignment as well as the high-resolution structure determination of polytopic membrane proteins in a detergent-free, near-native lipid bilayer setting. By applying this method to bacteriorhodopsin, we show that our smaller nanodiscs can also be beneficial for the structural characterization of the important class of seven-transmembrane helical proteins. Our set of engineered nanodiscs of subsequently smaller diameters can be used to screen for optimal NMR spectral quality for small to medium-sized membrane proteins while still providing a functional environment. In addition to their key improvements for de novo structure determination, due to their smaller size these nanodiscs enable the investigation of interactions between membrane proteins and their (soluble) partner proteins, unbiased by the presence of detergents that might disrupt biologically relevant interactions.
Authors:
Franz Hagn; Manuel Etzkorn; Thomas Raschle; Gerhard Wagner
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't     Date:  2013-01-25
Journal Detail:
Title:  Journal of the American Chemical Society     Volume:  135     ISSN:  1520-5126     ISO Abbreviation:  J. Am. Chem. Soc.     Publication Date:  2013 Feb 
Date Detail:
Created Date:  2013-02-06     Completed Date:  2013-08-01     Revised Date:  2014-02-13    
Medline Journal Info:
Nlm Unique ID:  7503056     Medline TA:  J Am Chem Soc     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1919-25     Citation Subset:  IM    
Data Bank Information
Bank Name/Acc. No.:
PDB/2M06;  2M07
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MeSH Terms
Descriptor/Qualifier:
Lipid Bilayers / chemistry*
Membrane Proteins / chemistry*
Models, Molecular
Nanostructures / chemistry*
Nuclear Magnetic Resonance, Biomolecular
Phospholipids / chemistry*
Protein Conformation
Grant Support
ID/Acronym/Agency:
GM047467/GM/NIGMS NIH HHS; GM094608/GM/NIGMS NIH HHS; P01 GM047467/GM/NIGMS NIH HHS; P01 GM62580/GM/NIGMS NIH HHS; P41-EB002026/EB/NIBIB NIH HHS; R01 GM075879/GM/NIGMS NIH HHS; S10 RR026417/RR/NCRR NIH HHS; S10 RR029236/RR/NCRR NIH HHS; U54 GM094608/GM/NIGMS NIH HHS
Chemical
Reg. No./Substance:
0/Lipid Bilayers; 0/Membrane Proteins; 0/Phospholipids
Comments/Corrections

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