title: Protein structural change at the cytoplasmic surface as the cause of cooperativity in the bacteriorhodopsin photocycle.
abstract: The effects of excitation light intensity on the kinetics of the bacteriorhodopsin photocycle were investigated. The earlier reported intensity-dependent changes at 410 and 570 nm are explained by parallel increases in two of the rate constants, for proton transfers to D96 from the Schiff base and from the cytoplasmic surface, without changes in the others, as the photoexcited fraction is increased. Thus, it appears that the pKa of D96 is raised by a cooperative effect within the purple membrane. This interpretation of the wild-type kinetics was confirmed by results with several mutant proteins, where the rates are well separated in time and a model-dependent analysis is unnecessary. Based on earlier results that demonstrated a structural change of the protein after deprotonation of the Schiff base that increases the area of the cytoplasmic surface, and the effects of high hydrostatic pressure and lowered water activity on the photocycle steps in question, we suggest that the pKa of D96 is raised by a lateral pressure that develops when other bacteriorhodopsin molecules are photoexcited within the two-dimensional lattice of the purple membrane. Expulsion of no more than a few water molecules bound near D96 by this pressure would account for the calculated increase of 0.6 units in the pKa.
authors.affiliation: Department of Physiology and Biophysics, University of California, Irvine 92717, USA.
authors.forename: G
authors.initials: G
authors.lastname: Váró
authors.forename: R
authors.initials: R
authors.lastname: Needleman
authors.forename: J K
authors.initials: JK
authors.lastname: Lanyi
mesh_terms: D001436:Bacteriorhodopsins / Q000737:chemistry* / Q000528:radiation effects*; D055592:Biophysical Phenomena; D001703:Biophysics; D003593:Cytoplasm / Q000737:chemistry; D006217:Halobacterium / Q000737:chemistry / Q000528:radiation effects; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D015394:Molecular Structure; D010777:Photochemistry; D013816:Thermodynamics
chemical_list: D001436:Bacteriorhodopsins