The photocycle and proton translocation pathway in a cyanobacterial ion-pumping rhodopsin.
The genome of thylakoidless cyanobacterium Gloeobacter violaceus encodes a fast-cycling rhodopsin capable of light-driven proton transport. We characterize the dark state, the photocycle, and the proton translocation pathway of GR spectroscopically. The dark state of GR contains predominantly all-trans-retinal and, similar to proteorhodopsin, does not show the light/dark adaptation. We found an unusually strong coupling between the conformation of the retinal and the site of Glu132, the homolog of Asp96 of BR. Although the photocycle of GR is similar to that of proteorhodopsin in general, it differs in accumulating two intermediates typical for BR, the L-like and the N-like states. The latter state has a deprotonated cytoplasmic proton donor and is spectrally distinct from the strongly red-shifted N intermediate known for proteorhodopsin. The proton uptake precedes the release and occurs during the transition to the O intermediate. The proton translocation pathway of GR is similar to those of other proton-pumping rhodopsins, involving homologs of BR Schiff base proton acceptor and donor Asp85 and Asp96 (Asp121 and Glu132). We assigned a pair of FTIR bands (positive at 1749 cm(-1) and negative at 1734 cm(-1)) to the protonation and deprotonation, respectively, of these carboxylic acids.
Miranda
Mylene R M
MR
Department of Physics, University of Guelph, Guelph, Ontario N1G 2W1, Canada.
Choi
Ah Rheum
AR
Shi
Lichi
L
Bezerra
Arandi G
AG
Jung
Kwang-Hwan
KH
Brown
Leonid S
LS
D000458:Cyanobacteria / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D007700:Kinetics; D008027:Light; D008081:Liposomes / Q000378:metabolism; D050505:Mutant Proteins / Q000737:chemistry / Q000378:metabolism; D009154:Mutation; D017494:Proton Pumps; D012243:Rhodopsin / Q000378:metabolism; D025604:Rhodopsins, Microbial / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015398:Signal Transduction; D017550:Spectroscopy, Fourier Transform Infrared; D013059:Spectrum Analysis, Raman
D008081:Liposomes; D050505:Mutant Proteins; D017494:Proton Pumps; D025604:Rhodopsins, Microbial; C413548:proteorhodopsin; D012243:Rhodopsin