TY - JOUR
T1 - Sensitivity and resolution enhancement of oriented solid-state NMR
T2 - Application to membrane proteins
AU - Gopinath, T.
AU - Mote, Kaustubh R.
AU - Veglia, Gianluigi
N1 - Funding Information:
This work is supported by the National Institute of Health (GM 64742 and 72701 to G.V.).
PY - 2013
Y1 - 2013
N2 - 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 15N 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 15N and 13C 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.
AB - 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 15N 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 15N and 13C 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.
KW - Chemical shift anisotropy
KW - Dipolar couplings
KW - Oriented solid-state NMR
KW - Resolution enhancement
KW - Sensitivity enhancement
UR - http://www.scopus.com/inward/record.url?scp=84887104891&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84887104891&partnerID=8YFLogxK
U2 - 10.1016/j.pnmrs.2013.07.004
DO - 10.1016/j.pnmrs.2013.07.004
M3 - Review article
C2 - 24160761
AN - SCOPUS:84887104891
VL - 75
SP - 50
EP - 68
JO - Progress in Nuclear Magnetic Resonance Spectroscopy
JF - Progress in Nuclear Magnetic Resonance Spectroscopy
SN - 0079-6565
ER -