Proton channel hydration and dynamics of a bacteriorhodopsin triple mutant with an M-state-like conformation.
The hydration and dynamics of purple membranes (PM) containing the bacteriorhodopsin (BR) triple mutant D96G/F171C/F219L were investigated by neutron diffraction coupled with H(2)O/D(2)O exchange and by energy-resolved neutron scattering. The mutant, which is active in proton transport (Tittor et al. in J. Mol. Biol. 319:555-565, 2002), has an "open" ground-state structure similar to that of the M intermediate in the photocycle of the wild type (wt) (Subramaniam and Henderson in Nature 406:653-657, 2000). The experiments demonstrated an increased proton channel hydration in the mutant PM compared with wt PM, in both high (86%) and low (57%) relative humidity. We suggest that this is due to the smaller side chains of the mutant residues liberating space for more water molecules in the proton channel, which would then be able to participate in the proton translocation network. PM thermal dynamics has been shown to be very sensitive to membrane hydration (Lehnert et al. in Biophys. J. 75:1945-1952, 1998). The global dynamical behaviour of the mutant PM on the 100-ps time scale, as a function of relative humidity, was found to be identical to that of the wt, showing that the "open" BR structure and additional water molecules in the proton channel do not provide a softer environment enabling increased flexibility.
Lehnert
U
U
Max-Planck-Institut für Biochemie, 82152 Martinsried, Germany.
Réat
V
V
Zaccai
G
G
Oesterhelt
D
D
D001436:Bacteriorhodopsins / Q000235:genetics*; D002462:Cell Membrane / Q000378:metabolism; D017666:Deuterium Oxide / Q000737:chemistry; D019617:Halobacterium salinarum / Q000378:metabolism; D006358:Hot Temperature; D008027:Light; D008055:Lipids / Q000737:chemistry; D008968:Molecular Conformation; D009154:Mutation; D009502:Neutrons; D011487:Protein Conformation; D011522:Protons; D018717:Purple Membrane / Q000378:metabolism; D012542:Scattering, Radiation; D013696:Temperature; D014867:Water / Q000737:chemistry
D008055:Lipids; D011522:Protons; D014867:Water; D001436:Bacteriorhodopsins; D017666:Deuterium Oxide