Fourier transform infrared evidence for proline structural changes during the bacteriorhodopsin photocycle.
Structural changes involving bacteriohodopsin proline residues have been investigated by Fourier transform infrared difference spectroscopy. Bacteriohodopsin (bR)-producing Halobacteria halobium were grown on a stringent medium containing either ring-perdeuterated proline or 15N-labeled proline. Comparison of the difference spectra obtained from the photoreactions of these labeled bR samples with those for unlabeled bR has led to the assignment of peaks due to proline vibrations. [proline-N15]bR exhibited a 15-cm-1 isotopic downshift of peaks in the 1420- to 1440-cm-1 region of the bR----K and bR----M difference spectra as well as a similar downshift of peaks found in the absolute absorption spectrum of bR. In contrast, [proline-D7]bR did not cause shifts in this region of the difference spectra. These results indicate that one or more prolines undergo a structural rearrangement during the bR photocycle involving the Xaa-Pro C--N peptide bond. This change may be directly coupled to the light-induced isomerization of the retinal chromophore from all-trans-retinal to 13-cis-retinal.
Rothschild
K J
KJ
Physics Department, Boston University, MA 02215.
He
Y W
YW
Gray
D
D
Roepe
P D
PD
Pelletier
S L
SL
Brown
R S
RS
Herzfeld
J
J
D001436:Bacteriorhodopsins / Q000378:metabolism* / Q000528:radiation effects; D005583:Fourier Analysis; D006217:Halobacterium / Q000378:metabolism; D007700:Kinetics; D011392:Proline; D011487:Protein Conformation; D013055:Spectrophotometry, Infrared / Q000379:methods
D001436:Bacteriorhodopsins; D011392:Proline