title: Detergent delipidation and solubilization strategies for high-resolution NMR of the membrane protein bacteriorhodopsin.
abstract: High-resolution NMR studies of bacteriorhodopsin require the availability of the detergent-solubilized protein with both high concentration and small rotational correlation time. A procedure is described for the optimized preparation of such samples. Bacteriorhodopsin was first delipidated by detergent treatment of purple membrane under nonsolubilizing conditions for the protein. The delipidated aggregated protein could then be solubilized into monomers at concentration close to millimolar by selected detergents. The solubilizing detergent had an important effect on the rotational correlation time of the protein as shown by measuring in each case the temperature-dependent stability of the protein, the size of the detergent-protein complex, and the detergent viscosity. Consistently, a strong influence of the detergent was also found on spectral resolution in 13C NMR spectra of solubilized bacteriorhodopsin labeled with [1-13C]phenylalanine. Best resolution was obtained using n-dodecylmaltoside as detergent, with which relatively narrow well resolved 13C NMR resonances were observed at 50 degrees C. It is suggested that high-resolution NMR studies performed with this detergent may contribute to the structural resolution of bacteriorhodopsin.
authors.affiliation: Unité de Recherches Associée au Centre National de la Recherche Scientifique 1290, Gif sur Yvette, France.
authors.forename: M
authors.initials: M
authors.lastname: Seigneuret
authors.forename: J M
authors.initials: JM
authors.lastname: Neumann
authors.forename: J L
authors.initials: JL
authors.lastname: Rigaud
mesh_terms: D001436:Bacteriorhodopsins / Q000737:chemistry*; D002793:Cholic Acids; D003902:Detergents; D009682:Magnetic Resonance Spectroscopy; D008563:Membrane Lipids / Q000737:chemistry*; D012995:Solubility
chemical_list: D002793:Cholic Acids; D003902:Detergents; D008563:Membrane Lipids; D001436:Bacteriorhodopsins; C028213:3-((3-cholamidopropyl)dimethylammonium)-1-propanesulfonate