Sensitivity and resolution enhancement of oriented solid-state NMR: application to membrane proteins. Prog Nucl Magn Reson Spectrosc 2013 Nov;75:50-68
Date
10/29/2013Pubmed ID
24160761Pubmed Central ID
PMC3850070DOI
10.1016/j.pnmrs.2013.07.004Scopus ID
2-s2.0-84887104891 (requires institutional sign-in at Scopus site) 31 CitationsAbstract
Oriented solid-state NMR (O-ssNMR) spectroscopy is a major technique for the high-resolution analysis of the structure and topology of transmembrane proteins in native-like environments. Unlike magic angle spinning (MAS) techniques, O-ssNMR spectroscopy requires membrane protein preparations that are uniformly oriented (mechanically or magnetically) so that anisotropic NMR parameters, such as dipolar and chemical shift interactions, can be measured to determine structure and orientation of membrane proteins in lipid bilayers. Traditional sample preparations involving mechanically aligned lipids often result in short relaxation times which broaden the (15)N resonances and encumber the manipulation of nuclear spin coherences. The introduction of lipid bicelles as membrane mimicking systems has changed this scenario, and the more favorable relaxation properties of membrane protein (15)N and (13)C resonances make it possible to develop new, more elaborate pulse sequences for higher spectral resolution and sensitivity. Here, we describe our recent progress in the optimization of O-ssNMR pulse sequences. We explain the theory behind these experiments, demonstrate their application to small and medium size proteins, and describe the technical details for setting up these new experiments on the new generation of NMR spectrometers.
Author List
Gopinath T, Mote KR, Veglia GAuthor
Gopinath Tata PhD Assistant Professor in the Biophysics department at Medical College of WisconsinMESH terms used to index this publication - Major topics in bold
AlgorithmsAnimals
Crystallization
Humans
Membrane Proteins
Nuclear Magnetic Resonance, Biomolecular
