title: The last phase of the reprotonation switch in bacteriorhodopsin: the transition between the M-type and the N-type protein conformation depends on hydration.
abstract: In order to elucidate the mechanism of the reprotonation switch of bacteriorhodopsin, the protein conformation of the M intermediate of the D96N mutant was examined at various hydration conditions by X-ray diffraction and FTIR spectroscopy. We observed two distinct protein conformations at different levels of hydration. One is like in the N photointermediate, although in this case with an unprotonated Schiff base. It is stabilized in highly hydrated samples. The other is a protein conformation identical to that in the normal M intermediate of wild-type bacteriorhodopsin, which is stabilized in partially dehydrated samples. The hydration dependence of the structural transition between the M-type and the N-type conformations suggests that there is a change in the binding of water at the cytoplasmic surface. Thus, more water molecules bind in the N-type structure than in the M-type. This is consistent with the idea that the conformational change from the M-type to the N-type corresponds to the opening of the proton channel to the cytoplasmic surface by tilt of the cytoplasmic end of helix F, and that this is required for proton transfer from Asp-96 to the retinal Schiff base.
authors.affiliation: Department of Physics, Graduate School of Science, Osaka University, Toyonaka, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Kamikubo
authors.forename: T
authors.initials: T
authors.lastname: Oka
authors.forename: Y
authors.initials: Y
authors.lastname: Imamoto
authors.forename: F
authors.initials: F
authors.lastname: Tokunaga
authors.forename: J K
authors.initials: JK
authors.lastname: Lanyi
authors.forename: M
authors.initials: M
authors.lastname: Kataoka
mesh_terms: D001216:Asparagine / Q000235:genetics; D001224:Aspartic Acid / Q000235:genetics; D001436:Bacteriorhodopsins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D016254:Mutagenesis, Insertional; D011487:Protein Conformation; D011522:Protons; D012545:Schiff Bases; D017550:Spectroscopy, Fourier Transform Infrared; D014867:Water; D014961:X-Ray Diffraction
chemical_list: D011522:Protons; D012545:Schiff Bases; D014867:Water; D001224:Aspartic Acid; D001436:Bacteriorhodopsins; D001216:Asparagine