Advanced solid-state NMR techniques for characterization of membrane protein structure and dynamics: Application to Anabaena Sensory Rhodopsin

Biosilica and bioinspired silica studied by solid-state NMR

“…[98][99][100][101][102][103][104] The physical condition of a sample is a key determinant for NMR resolution, with crystalline preparation usually yielding more resolved spectra [105]. However, the most interesting targets for solid state NMR are precisely those that are harder (or impossible) to obtain in crystalline form, such as membrane proteins, [5][6][7][106][107][108][109] protein assemblies [110][111][112][113][114], pathological aggregates [57][58][59][60] and fibrils [119][120][121][122][123]3,[124][125][126].…”

Recent Solid-State NMR Studies of Membrane-Bound Peptides and Proteins

“…A number of methods have since been developed that permit the selective reintroduction of these anisotropic interactions to facilitate their analysis. These methods and their applications to biological systems have been extensively reviewed [10,11].…”

“…The extended conformation of the C-terminal Lys-Leu-Lys-OH segment with peptide bonds (HdNdC]O dipoles) perpendicular to the membrane surface was chosen for modeling. In this structural model, the message segment, Dyn(1-4), contacts hydrophobic membrane layers, and the address segment, Dyn (5)(6)(7)(8)(9)(10)(11)(12)(13), is in contact with the aqueous phase. The results with IR-ATR spectroscopy analyses indicated that a strong increase in helicity occurs upon contact with neutral lecithin membranes.…”

“…These experiments enable the correlation of 13 C and 15 N signals for assignment to amino acid residues and chemical shifts and distance constraints. These data can then be used to determine the high-resolution structures of membrane-associated peptides and proteins [10,11].…”

Isotope Labeling of Eukaryotic Membrane Proteins in Yeast for Solid-State NMR

“…Besides the main technical challenges related to functional expression, stability in detergents, and crystallization, X-ray crystallography and solution NMR often face a conceptual problem of providing a native-like environment for membrane proteins, being restricted to using three-dimensional protein crystals or small detergent micelles. In this respect, an emerging powerful technique of solid-state NMR (ssNMR) provides an attractive alternative, as it allows studies of membrane proteins in the artificial native-like lipid bilayers or even in the native membranes, without having any inherent limitations on the protein molecular weight (Hong, Zhang, & Hu, 2012;Judge, Taylor, Dannatt, & Watts, 2015;Murray, Das, & Cross, 2013;Opella, 2015;Renault et al, 2012;Wang & Ladizhansky, 2014;Ward, Brown, & Ladizhansky, 2015). In the last few years, several structures of polytopic membrane proteins and their oligomers have been determined by ssNMR (Das et al, 2015;Park et al, 2012;Shahid et al, 2012;Wang et al, 2013), making it a reputable player in the team of structural biology techniques.…”