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Structure determination of membrane proteins in five easy pieces. Methods 2011 Dec;55(4):363-9

Date

10/04/2011

Pubmed ID

21964394

Pubmed Central ID

PMC3264820

DOI

10.1016/j.ymeth.2011.09.009

Scopus ID

2-s2.0-84855993501 (requires institutional sign-in at Scopus site)   26 Citations

Abstract

Rotational Alignment (RA) solid-state NMR provides the basis for a general method for determining the structures of membrane proteins in phospholipid bilayers under physiological conditions. Membrane proteins are high priority targets for structure determination, and are challenging for existing experimental methods. Because membrane proteins reside in liquid crystalline phospholipid bilayer membranes it is important to study them in this type of environment. The RA solid-state NMR approach we have developed can be summarized in five steps, and incorporates methods of molecular biology, biochemistry, sample preparation, the implementation of NMR experiments, and structure calculations. It relies on solid-state NMR spectroscopy to obtain high-resolution spectra and residue-specific structural restraints for membrane proteins that undergo rotational diffusion around the membrane normal, but whose mobility is otherwise restricted by interactions with the membrane phospholipids. High resolution spectra of membrane proteins alone and in complex with other proteins and ligands set the stage for structure determination and functional studies of these proteins in their native, functional environment.

Author List

Marassi FM, Das BB, Lu GJ, Nothnagel HJ, Park SH, Son WS, Tian Y, Opella SJ

Author

Francesca M. Marassi PhD Chair, Professor in the Biophysics department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Animals
Bacterial Proteins
Cation Transport Proteins
Humans
Liposomes
Membrane Proteins
Nuclear Magnetic Resonance, Biomolecular
Protein Structure, Tertiary
Receptors, Interleukin-8A