title: Light-induced isomerization causes an increase in the chromophore tilt in the M intermediate of bacteriorhodopsin: a neutron diffraction study.
abstract: Bacteriorhodopsin (BR) was regenerated with two selectively deuterated retinals, one with 11 deuterons in the beta-ionone ring (D11) and the other with 5 deuterons (D5) at the end of the polyene chain closest to the Schiff base at carbon atoms C-14, C-15, and C-20. Both label positions (centers of deuteration) were obtained from difference Fourier maps of projections onto the plane of the membrane by neutron diffraction at 90 K, both in the light-adapted ground-state BR568 and in the photocycle intermediate M412. To retard the decay of M412, purple membrane films were soaked in 0.1 M or 1 M guanidine hydrochloride at pH 9.6. M412 was produced by illuminating oriented membrane films at physiological temperature (278 K), followed by rapid cooling to 90 K in the absence of light. The results show that in the projected structure the ring position is unaltered during the transition from BR568 to M412, whereas the position of the D5 label shifts by 1.4 +/- 0.9 A toward the ring. The shortened interlabel distance in the projected structure for the M412 state implies that as a result of the all-trans/13-cis isomerization, the C-5 to C-13 part of the polyene chain tilts out of the plane of the membrane toward the cytoplasm by about 11 degrees +/- 6 degrees. Pairwise comparison of data sets with the same retinal for the two photocycle states M412 and BR568 leads to four difference-density maps for the protein, which are in agreement with previous work. They show changes in the protein density near helices G and F.
authors.affiliation: Department of Physics, Freie Universität Berlin, Germany.
authors.forename: T
authors.initials: T
authors.lastname: Hauss
authors.forename: G
authors.initials: G
authors.lastname: Büldt
authors.forename: M P
authors.initials: MP
authors.lastname: Heyn
authors.forename: N A
authors.initials: NA
authors.lastname: Dencher
mesh_terms: D001436:Bacteriorhodopsins / Q000737:chemistry* / Q000528:radiation effects; D005583:Fourier Analysis; D006217:Halobacterium / Q000378:metabolism; D007536:Isomerism; D008027:Light; D009502:Neutrons; D011487:Protein Conformation; D013816:Thermodynamics
chemical_list: D001436:Bacteriorhodopsins