NMR of membrane proteins in micelles and bilayers: the FXYD family proteins. Methods 2007 Apr;41(4):398-408
Date
03/21/2007Pubmed ID
17367712Pubmed Central ID
PMC2920895DOI
10.1016/j.ymeth.2006.08.011Scopus ID
2-s2.0-33947171594 (requires institutional sign-in at Scopus site) 37 CitationsAbstract
Determining the atomic resolution structures of membrane proteins is of particular interest in contemporary structural biology. Helical membrane proteins constitute one-third of the expressed proteins encoded in a genome, many drugs have membrane-bound proteins as their receptors, and mutations in membrane proteins result in human diseases. Although integral membrane proteins provide daunting technical challenges for all methods of protein structure determination, nuclear magnetic resonance (NMR) spectroscopy can be an extremely versatile and powerful method for determining their structures and characterizing their dynamics, in lipid environments that closely mimic the cell membranes. Once milligram amounts of isotopically labeled protein are expressed and purified, micelle samples can be prepared for solution NMR analysis, and lipid bilayer samples can be prepared for solid-state NMR analysis. The two approaches are complementary and can provide detailed structural and dynamic information. This paper describes the steps for membrane protein structure determination using solution and solid-state NMR. The methods for protein expression and purification, sample preparation and NMR experiments are described and illustrated with examples from the FXYD proteins, a family of regulatory subunits of the Na,K-ATPase.
Author List
Franzin CM, Gong XM, Thai K, Yu J, Marassi FMAuthor
Francesca M. Marassi PhD Chair, Professor in the Biophysics department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
Amino Acid SequenceCell Line, Transformed
Detergents
Gene Expression
Intracellular Signaling Peptides and Proteins
Lipid Bilayers
Membrane Proteins
Micelles
Molecular Sequence Data
Neoplasm Proteins
Nitrogen Isotopes
Nuclear Magnetic Resonance, Biomolecular
Phosphoproteins
Phosphorus Isotopes
Plasmids
Potassium Channels
Protein Structure, Tertiary
Sodium-Potassium-Exchanging ATPase
