fd coat protein structure in membrane environments: structural dynamics of the loop between the hydrophobic trans-membrane helix and the amphipathic in-plane helix

Complete resolution of the solid-state NMR spectrum of a uniformly 15 N-labeled membrane protein in phospholipid bilayers

“…The observed ratios are about 10, 30, and 50 for the 1 H chemical shift, 1 H-15 N dipolar coupling, and 15 N chemical shift dimensions, respectively. These ratios are comparable to those observed along the 1 H and 15 N frequency axes of multidimensional solution NMR spectra of the same protein in detergent micelles (32); even though the linewidths are considerably narrower in solution NMR spectra, the range of isotropic chemical shift frequencies are correspondingly smaller as well. Note that three independent parameters contribute to the resolution in three-dimensional solid-state NMR correlation spectra compared with only two in typical three-dimensional solution NMR spectra, for example NOESY-HMQC spectra (11,12).…”

“…Typically, high quality two-dimensional spectra are obtained in overnight runs, whereas the three-dimensional data shown in Fig. 3 were obtained in 3 days, the same length of time as the corresponding solution nuclear Overhauser effect spectroscopyheteronuclear multiple quantum correlation (NOESY-HMQC) spectrum from a sample in micelles (32).…”

“…Newly synthesized copies of the protein are stored in the membrane of infected Escherichia coli bacteria prior to incorporation into virus particles as they are extruded through the membrane. The structure of the membrane bound form of this protein has been determined in micelles by solution NMR spectroscopy (29)(30)(31)(32). It has an amphipathic helix that lies in the plane of the membrane (residues 7-16), and a longer hydrophobic membrane spanning helix (residues 27-44).…”

Positively and negatively charged residues have different effects on the position in the membrane of a model transmembrane helix

“…In order to extend our set of reference TMHs and, in particular, in order to determine the MGD for a TMH with known position in a bilayer and containing charged residues near its C-terminal end, we chose to study the TMH from the phage M13 major coat protein. This TMH has been shown by NMR to extend from approximately residue Ala25 to Phe45 (Almeida & Opella, 1997). Further studies in detergents and model lipid bilayers have con®rmed this structure, and have located Ala25 and Thr46 in a somewhat hydrophobic environment close to the membrane/water interface Stopar et al, 1996).…”

Application of solid-state NMR restraint potentials in membrane protein modeling

“…The structure was determined by solution NMR (PDB:1FDM) [31] as well as SSNMR (PDB:1MZT) [12]. Following the structure determination protocol, we obtained 45 (vacuum) and 61 (GBSW) structures that satisfy the selection criteria (δσ ≤ 1.0 ppm and δν ≤ 1.0 kHz).…”