Phototransformation and proton pumping activity of the 14-fluoro bacteriorhodopsin derivatives.
The photoinduced behavior and proton pumping characteristics of some bacteriorhodopsin (BR) analogs with fluorinated chromophores (all-trans 14-fluorinated [14-F] retinal and 13-cis 14-F retinal) derived from wild type (WT) and D96N mutant BR were investigated. These analogs were characterized using spectrophotometry and a highly sensitive electrochemical technique. Similar to the white membrane JW2N, the apomembranes WT ET 1000 and D96N form photoactive pigments with the 14-F chromophores. The resulting analogs have a major absorption band at 588 nm. Red-shifted pigment (lambdamax</=680 nm) has been previously observed as a minor component of the major 587-nm pigment in 14-F BR made with white membrane JW2N. A similar red-shifted pigment is formed under yellow light (lambda>500 nm) only in the 14-F analogs derived from WT ET 1000. The measurements of the photoinduced transformation in 14-F WT analogs show that the photocycle of the major pigment occurs simultaneously with the process in the red region and is partially masked by the formation of the red-shifted species. The 14-F D96N samples have a significantly slower and more complicated photoinduced behavior. Electrochemical measurements show that the photoinduced transformation of the red species is not accompanied by proton transport.
Druzhko
A B
AB
Biotechnology Division, National Institute of Standards and Technology, Gaithersburg, MD 20899, USA. druzhko@venus.iteb.serpukhov.su
Robertson
B
B
Alvarez
R
R
de Lera
A R
AR
Weetall
H H
HH
D001436:Bacteriorhodopsins / Q000031:analogs & derivatives* / Q000737:chemistry* / Q000528:radiation effects; D019617:Halobacterium salinarum; D006863:Hydrogen-Ion Concentration; D007536:Isomerism; D007700:Kinetics; D008027:Light; D010777:Photochemistry; D017494:Proton Pumps / Q000528:radiation effects; D012172:Retinaldehyde / Q000031:analogs & derivatives* / Q000737:chemistry / Q000528:radiation effects; D013056:Spectrophotometry, Ultraviolet
D017494:Proton Pumps; D001436:Bacteriorhodopsins; C065196:14-fluororetinal; D012172:Retinaldehyde