Distortions in the photocycle of bacteriorhodopsin at moderate dehydration.
The photoreaction of bacteriorhodopsin was studied in moderately dehydrated films (relative humidities between 100 and 65%). Time-resolved difference spectra from a gated optical multichannel analyzer, between 100 ns and 100 ms after photoexcitation, were decomposed into sums of difference spectra of the intermediates K, L, M, N, and O, and the kinetics obtained were fitted to various alternative schemes. The data confirm the model of a single reaction sequence with reversible reactions we proposed recently for purple membrane suspensions (Váró, G., and J. K. Lanyi. Biochemistry. 1990. 29:2241-2250) but including reversibility also for the reaction K in equilibrium with L in addition to L in equilibrium with M, M in equilibrium with N, and N in equilibrium with O. With increasing dehydration the kinetics were increasingly dominated by the reverse reactions. As before, fitting the data required the existence of two M species in series: L in equilibrium with M1 in equilibrium with M2 in equilibrium with N. The M1 in equilibrium with M2 reaction was greatly slowed at lower humidities. This step might be the switch for the unidirectional transfer of protons. With increasing dehydration recovery of BR occurred less and less via the N intermediate and increasingly via direct shunts from the two M species. As indicated earlier by electrical measurements with similarly dried bacteriorhodopsin films (Váró, G., and L. Keszthelyi, 1983. Biophys. J. 43:47-51). The latter are pathways not necessarily associated with net proton translocation.
Váró
G
G
Department of Physiology and Biophysics, University of California, Irvine 92717.
Lanyi
J K
JK
D001436:Bacteriorhodopsins / Q000737:chemistry / Q000378:metabolism* / Q000528:radiation effects; D006217:Halobacterium / Q000378:metabolism; D007700:Kinetics; D008962:Models, Theoretical; D010777:Photochemistry; D013053:Spectrophotometry
D001436:Bacteriorhodopsins