Internal molecular motions of bacteriorhodopsin: hydration-induced flexibility studied by quasielastic incoherent neutron scattering using oriented purple membranes.
Quasielastic incoherent neutron scattering from hydrogen atoms, which are distributed nearly homogeneously in biological molecules, allows the investigation of diffusive motions occurring on the pico- to nanosecond time scale. A quasielastic incoherent neutron scattering study was performed on the integral membrane protein bacteriorhodopsin (BR), which is a light-driven proton pump in Halobacterium salinarium. BR is embedded in lipids, forming patches in the cell membrane of the organism, which are the so called purple membranes (PMs). Measurements were carried out at room temperature on oriented PM-stacks hydrated at two different levels (low hydration, h = 0.03 g of D2O per g of PM; high hydration, h = 0.28 g of D2O per g of PM) using time-of-flight spectrometers. From the measured spectra, different diffusive components were identified and analyzed with respect to the influence of hydration. This study supports the idea that a decrease in hydration results in an appreciable decrease in internal molecular flexibility of the protein structure. Because it is known from studies on the function of BR that the pump activity is reduced if the hydration level of the protein is insufficient, we conclude that the observed diffusive motions are essential for the function of this protein. A detailed analysis and classification of the different kinds of diffusive motions, predominantly occurring in PMs under physiological conditions, is presented.
Fitter
J
J
Hahn-Meitner-Institut, Berlin Neutron Scattering Center, Germany.
Lechner
R E
RE
Buldt
G
G
Dencher
N A
NA
D001436:Bacteriorhodopsins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D017666:Deuterium Oxide; D004058:Diffusion; D004548:Elasticity; D006217:Halobacterium; D007700:Kinetics; D008956:Models, Chemical; D009502:Neutrons; D018717:Purple Membrane / Q000737:chemistry*; D012542:Scattering, Radiation
D001436:Bacteriorhodopsins; D017666:Deuterium Oxide