title: Vibrational spectroscopy of bacteriorhodopsin mutants: light-driven proton transport involves protonation changes of aspartic acid residues 85, 96, and 212.
abstract: Fourier transform infrared (FTIR) difference spectra have been obtained for the bR----K, bR----L, and bR----M photoreactions in bacteriorhodopsin mutants in which Asp residues 85, 96, 115, and 212 have been replaced by Asn and by Glu. Difference peaks that had previously been attributed to Asp COOH groups on the basis of isotopic labeling were absent or shifted in these mutants. In general, each COOH peak was affected strongly by mutation at only one of the four residues. Thus, it was possible to assign each peak tentatively to a particular Asp. From these assignments, a model for the proton-pumping mechanism of bR is derived, which features proton transfers among Asp-85, -96, and -212, the chromophore Schiff base, and other ionizable groups within the protein. The model can explain the observed COOH peaks in the FTIR difference spectra of bR photointermediates and could also account for other recent results on site-directed mutants of bR.
authors.affiliation: Physics Department and Program in Cellular Biophysics, Boston University, Massachusetts 02215.
authors.forename: M S
authors.initials: MS
authors.lastname: Braiman
authors.forename: T
authors.initials: T
authors.lastname: Mogi
authors.forename: T
authors.initials: T
authors.lastname: Marti
authors.forename: L J
authors.initials: LJ
authors.lastname: Stern
authors.forename: H G
authors.initials: HG
authors.lastname: Khorana
authors.forename: K J
authors.initials: KJ
authors.lastname: Rothschild
mesh_terms: D001224:Aspartic Acid; D001436:Bacteriorhodopsins / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D005583:Fourier Analysis; D005796:Genes; D005798:Genes, Bacterial; D006217:Halobacterium / Q000235:genetics / Q000378:metabolism; D008027:Light; D009154:Mutation; D011522:Protons; D013055:Spectrophotometry, Infrared
chemical_list: D011522:Protons; D001224:Aspartic Acid; D001436:Bacteriorhodopsins