title: Towards high-resolution solid-state NMR on large uniformly 15N- and [13C,15N]-labeled membrane proteins in oriented lipid bilayers.
abstract: Based on exact numerical simulations, taking into account isotropic and conformation-dependent anisotropic nuclear spin interactions, we systematically analyse the prospects for high-resolution solid-state NMR on large isotope-labeled membrane proteins macroscopically oriented in phospholipid bilayers. Using the known X-ray structures of rhodopsin and porin as models for large membrane proteins with typical alpha-helical and beta-barrel structural motifs, the analysis considers all possible one- to six-dimensional spectra comprised of frequency dimensions with evolution under any combination of amide 1H, amide 15N, and carbonyl 13C chemical shifts as well as 1H-15N dipole-dipole couplings. Under consideration of typical nuclear spin interaction and experimental line-shape parameters, the analysis provides new insight into the resolution capability and orientation-dependent transfer efficiency of existing experiments as well as guidelines as to improved experimental approaches for the study of large uniformly 15N- and [13C,15N]-labeled membrane proteins. On basis of these results and numerical optimizations of coherence-transfer efficiencies, we propose several new high-resolution experiments for sequential protein backbone assignment and structure determination.
authors: Department of Molecular and Structural Biology, University of Aarhus, Denmark. tv@imsb.av.dk
authors: Thomas
authors: T
authors: Vosegaard
authors: Niels Chr
authors: NC
authors: Nielsen
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D002247:Carbon Isotopes; D008051:Lipid Bilayers / Q000737:chemistry*; D008563:Membrane Lipids / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry*; D008958:Models, Molecular; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular / Q000379:methods*; D010743:Phospholipids / Q000737:chemistry; D018272:Porins / Q000737:chemistry; D011487:Protein Conformation; D012107:Research Design; D012243:Rhodopsin / Q000737:chemistry; D012680:Sensitivity and Specificity; D012984:Software
chemical_list: D002247:Carbon Isotopes; D008051:Lipid Bilayers; D008563:Membrane Lipids; D008565:Membrane Proteins; D009587:Nitrogen Isotopes; D010743:Phospholipids; D018272:Porins; D012243:Rhodopsin