Helix conformations in 7TM membrane proteins determined using oriented-sample solid-state NMR with multiple residue-specific 15N labeling.

Journal article

Oriented solid-state NMR in combination with multiple-residue-specific (15)N labeling and extensive numerical spectral analysis is proposed to determine helix conformations of large membrane proteins in native membranes. The method is demonstrated on uniaxially oriented samples of (15)N-methionine, -valine, and -glycine-labeled bacteriorhopsin in native purple membranes. Experimental two-dimensional (1)H-(15)N dipole-dipole coupling versus (15)N chemical shift spectra for all samples are analyzed numerically to establish combined constraints on the orientation of the seven transmembrane helices relative to the membrane bilayer normal. Since the method does not depend on specific resonance assignments and proves robust toward nonidealities in the sample alignment, it may be generally feasible for the study of conformational arrangement and function-induced conformation changes of large integral membrane proteins.

Authors: Vosegaard, T; Kamihira-Ishijima, M; Watts, A; Nielsen, N

• Publication status: Published
• Journal: Biophysical journal
• Volume: 94
• Issue: 1
• Pages: 241-250
• Publication date: 2008-01-01
• DOI: 10.1529/biophysj.107.116004
• EISSN: 1542-0086
• ISSN: 0006-3495
• Language: English
• Keywords: Nitrogen Isotopes; Protein Conformation; Magnetic Resonance Spectroscopy; Models, Chemical; Adenosine Triphosphatases; Models, Molecular; Amino Acids; Membrane Proteins; Computer Simulation