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Conformational heterogeneity of transmembrane residues after the Schiff base reprotonation of bacteriorhodopsin
Author(s) -
Mason A. James,
Turner George J.,
Glaubitz Clemens
Publication year - 2005
Publication title -
the febs journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.981
H-Index - 204
eISSN - 1742-4658
pISSN - 1742-464X
DOI - 10.1111/j.1742-4658.2005.04633.x
Subject(s) - bacteriorhodopsin , chemistry , schiff base , magic angle spinning , nuclear magnetic resonance spectroscopy , quenching (fluorescence) , crystallography , halobacterium , resonance (particle physics) , analytical chemistry (journal) , membrane , stereochemistry , biochemistry , chromatography , fluorescence , physics , particle physics , quantum mechanics
bR, N‐like and O‐like intermediate states of [ 15 N]methionine‐labelled wild type and D85N/T170C bacteriorhodopsin were accumulated in native membranes by controlling the pH of the preparations. 15 N cross polarization and magic angle sample spinning (CPMAS) NMR spectroscopy allowed resolution of seven out of nine resonances in the bR‐state. It was possible to assign some of the observed resonances by using 13 C/ 15 N rotational echo double resonance (REDOR) NMR and Mn 2+ quenching as well as D 2 O exchange, which helps to identify conformational changes after the bacteriorhodopsin Schiff base reprotonation. The significant differences in chemical shifts and linewidths detected for some of the resonances in N‐ and O‐like samples indicate changes in conformation, structural heterogeneity or altered molecular dynamics in parts of the protein.