The effects of octanol on the late photointermediates of rhodopsin.
Membrane suspensions of unperturbed rhodopsin and rhodopsin perturbed with 2.5 mM octanol were photolyzed with 477 nm laser pulses at 20 degrees C and 35 degrees C. Changes in absorbance were monitored at times ranging from 1 microsecond to 80 ms after excitation. The data were analyzed using singular value decomposition, global exponential fitting and kinetic modeling. A recently proposed model involving the photointermediate Meta-I380 (T. E. Thorgeirsson, J. W. Lewis, S. E. Wallace-Williams, and D. S. Kliger, Biochemistry 32, 13861-13872, 1993) fits data for samples with and without octanol. Comparison of the microscopic rates shows this alcohol accelerates the formation of Meta-II via Meta-I380. Activation and equilibrium thermodynamic parameters obtained from Arrhenius plots suggest that octanol reduces the entropy increase in forming both Meta-I380 and Meta-II. It also lowers the enthalpy of Meta-I380 relative to Lumi and of Meta-II relative to Meta-I480. To help determine whether octanol affects the protein directly or indirectly through the lipid bilayer, similar experiments were conducted using rhodopsin solubilized in 0.13% dodecyl maltoside with and without octanol. Spectral shifts in the presence of octanol suggest that a direct protein interaction exists in addition to previously reported effects dependent on membrane free volume.
Mah
T L
TL
Department of Chemistry and Biochemistry, University of California, Santa Cruz 95064, USA.
Szundi
I
I
Lewis
J W
JW
Jäger
S
S
Kliger
D S
DS
D000818:Animals; D002417:Cattle; D007700:Kinetics; D007834:Lasers; D008027:Light; D000442:Octanols / Q000494:pharmacology*; D010782:Photolysis; D012243:Rhodopsin / Q000737:chemistry* / Q000187:drug effects / Q000528:radiation effects; D012374:Rod Cell Outer Segment / Q000378:metabolism; D013053:Spectrophotometry
D000442:Octanols; D012243:Rhodopsin