Halide dependence of the halorhodopsin photocycle as measured by time-resolved infrared spectra.
Time-resolved Fourier transform infrared (FTIR) difference spectra of the halorhodopsin (hR) photocycle have been collected from 3 micros to 100 ms in saturating concentrations of KCl or KBr. Kinetic analysis of these data revealed two decay processes, with time constants of tau(1) approximately 150 micros and tau(2) approximately 16 ms in the presence of either halide, with tau(2) describing the return to the starting (hR) state. Comparison to previous low-temperature FTIR spectra of hR intermediates confirms that characteristic hK and hL spectral features are both present before the tau(1) decay, in a state previously defined as hK(L) (Dioumaev, A., and M. Braiman. 1997. Photochem. Photobiol. 66:755-763). However, the relative sizes of these features depend on which halide is present. In Br-, the hL features are clearly more dominant than in Cl-. Therefore, the state present before tau(1) is probably best described as an hK(L)/hL(1) equilibrium, instead of a single hK(L) state. Different halides affect the relative amounts of hK(L) and hL(1) present, i.e., Cl- produces a much more significant back-reaction from hL(1) to hK(L) than does Br-. The halide dependence of this back-reaction could therefore explain the halide selectivity of the halorhodopsin anion pump.
Hutson
M S
MS
Biophysics Program, University of Virginia Health Sciences Center 456, Charlottesville, Virginia 22908, USA.
Shilov
S V
SV
Krebs
R
R
Braiman
M S
MS
D001436:Bacteriorhodopsins / Q000737:chemistry* / Q000187:drug effects / Q000502:physiology; D001965:Bromides / Q000494:pharmacology*; D019617:Halobacterium salinarum / Q000502:physiology; D025602:Halorhodopsins; D007700:Kinetics; D008027:Light; D011189:Potassium Chloride / Q000494:pharmacology*; D017680:Potassium Compounds / Q000494:pharmacology*; D012172:Retinaldehyde / Q000737:chemistry / Q000378:metabolism; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods; D013816:Thermodynamics; D013997:Time Factors
D001965:Bromides; D025602:Halorhodopsins; D017680:Potassium Compounds; D001436:Bacteriorhodopsins; D011189:Potassium Chloride; C039004:potassium bromide; D012172:Retinaldehyde