The tertiary structural changes in bacteriorhodopsin occur between M states: X-ray diffraction and Fourier transform infrared spectroscopy.
The tertiary structural changes occurring during the photocycle of bacteriorhodopsin (BR) are assigned by X-ray diffraction to distinct M states, M1 and M2. Purple membranes (PM) of the mutant Asp96Asn at 15, 57, 75 and 100% relative humidity (r.h.) were studied in a parallel X-ray diffraction and Fourier transform infrared (FTIR) spectroscopic investigation. Light-dependent conformational changes of BR-Asp96Asn are observed at high hydration levels (100 and 75% r.h.) but not in partially dehydrated samples (57 and 15% r.h.). The FTIR spectra of continuously illuminated samples at low and high hydration, despite some differences, are characteristic of the M intermediate. The changes in diffraction patterns of samples in the M2 state are of the same magnitude as those of wild-type samples trapped with GuaHCl in the M(G) state. Additional large changes in the amide bands of the FTIR spectra occur between M2 and M(G). This suggests, that the tertiary structural changes between M1 and M2 are responsible for the switch opening the cytoplasmic half-channel of BR for reprotonation to complete the catalytic cycle. These tertiary structural changes seem to be triggered by a charge redistribution which might be a common feature of retinal proteins also in signal transduction.
Sass
H J
HJ
Forschungszentrum Jülich, IBI-2: Structural Biology, Germany. Jurgen@ibistr.dnet.kfa-juelich.de
Schachowa
I W
IW
Rapp
G
G
Koch
M H
MH
Oesterhelt
D
D
Dencher
N A
NA
Büldt
G
G
D000595:Amino Acid Sequence; D001216:Asparagine; D001224:Aspartic Acid; D001436:Bacteriorhodopsins / Q000737:chemistry* / Q000378:metabolism; D002384:Catalysis; D003624:Darkness; D019791:Guanidine; D006146:Guanidines; D006217:Halobacterium / Q000378:metabolism; D008027:Light; D017354:Point Mutation; D011489:Protein Denaturation; D017434:Protein Structure, Tertiary; D017550:Spectroscopy, Fourier Transform Infrared; D014961:X-Ray Diffraction
D006146:Guanidines; D001224:Aspartic Acid; D001436:Bacteriorhodopsins; D001216:Asparagine; D019791:Guanidine