Isotope labeling for solution and solid-state NMR spectroscopy of membrane proteins.
In this chapter, we summarize the isotopic labeling strategies used to obtain high-quality solution and solid-state NMR spectra of biological samples, with emphasis on integral membrane proteins (IMPs). While solution NMR is used to study IMPs under fast tumbling conditions, such as in the presence of detergent micelles or isotropic bicelles, solid-state NMR is used to study the structure and orientation of IMPs in lipid vesicles and bilayers. In spite of the tremendous progress in biomolecular NMR spectroscopy, the homogeneity and overall quality of the sample is still a substantial obstacle to overcome. Isotopic labeling is a major avenue to simplify overlapped spectra by either diluting the NMR active nuclei or allowing the resonances to be separated in multiple dimensions. In the following we will discuss isotopic labeling approaches that have been successfully used in the study of IMPs by solution and solid-state NMR spectroscopy.
Department of Biochemistry, University of Minnesota, Minneapolis, MN 55455, USA.
Raffaello
R
Verardi
Nathaniel J
NJ
Traaseth
Larry R
LR
Masterson
Vitaly V
VV
Vostrikov
Gianluigi
G
Veglia
D007553:Isotope Labeling / Q000379:methods*; D008565:Membrane Proteins / Q000737:chemistry* / Q000302:isolation & purification; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*
D008565:Membrane Proteins