Investigation of the primary photochemistry of bacteriorhodopsin by low-temperature Fourier-transform infrared spectroscopy.
The method of Fourier-transform infrared difference spectroscopy was applied to investigate the transition at 77K of bacteriorhodopsin in its light-adapted form to K6(10), the first intermediate which is stable at low temperature. In addition to unmodified bacteriorhodopsin, bacteriorhodopsin in 2H2O and bacteriorhodopsin containing [15-2H]retinal was used. The results show that major rearrangements occur in the Schiff base in this transition. It is not possible to identify a C = N stretching vibration of the Schiff base in K6(10). The identification of an N-H bending vibration in K6(10) shows that the nitrogen of the previous Schiff base still has a proton attached. The fingerprint region exhibits very unusual features for K6(10) and bears no similarity to protonated retinylidene Schiff base model compounds of any isomeric composition. Therefore, no conclusions on the isomeric state of the retinal in K6(10) can be drawn. The spectra show that the terminal part of the retinal is predominantly reflected in the difference spectra. This indicates that the most polar part of the retinal is located near the Schiff base. We have evidence for protein molecular changes occurring in this transition at 77K.
D001436:Bacteriorhodopsins / Q000302:isolation & purification*; D002338:Carotenoids / Q000302:isolation & purification*; D005583:Fourier Analysis; D006217:Halobacterium / Q000032:analysis; D008027:Light; D008565:Membrane Proteins / Q000302:isolation & purification*; D010777:Photochemistry; D013055:Spectrophotometry, Infrared / Q000379:methods; D013696:Temperature
D008565:Membrane Proteins; D002338:Carotenoids; D001436:Bacteriorhodopsins