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@ARTICLE{Efremov:52270,
author = {Efremov, R. and Gordeliy, V. I. and Heberle, J. and Büldt,
G.},
title = {{T}ime-resolved microscopy on a single crystal of
bacteriorhodopsin reveals lattice induced differences in the
photocycle kinetics},
journal = {Biophysical journal},
volume = {91},
issn = {0006-3495},
address = {New York, NY},
publisher = {Rockefeller Univ. Press},
reportid = {PreJuSER-52270},
pages = {1441 - 1451},
year = {2006},
note = {Record converted from VDB: 12.11.2012},
abstract = {The determination of the intermediate state structures of
the bacteriorhodopsin photocycle has lead to an
unprecedented level of understanding of the catalytic
process exerted by a membrane protein. However, the
crystallographic structures of the intermediate states are
only relevant if the working cycle is not impaired by the
crystal lattice. Therefore, we applied visible and Fourier
transform infrared spectroscopy (FTIR) microspectroscopy
with microsecond time resolution to compare the
photoreaction of a single bacteriorhodopsin crystal to that
of bacteriorhodopsin residing in the native purple membrane.
The analysis of the FTIR difference spectra of the resolved
intermediate states reveals great similarity in structural
changes taking place in the crystal and in PM. However, the
kinetics of the photocycle are significantly altered in the
three-dimensional crystal as compared to PM. Strikingly, the
L state decay is accelerated in the crystal, whereas the M
decay is delayed. The physical origin of this deviation and
the implications for trapping of intermediate states are
discussed. As a methodological advance, time-resolved
step-scan FTIR spectroscopy on a single protein crystal is
demonstrated for the first time which may be used in the
future to gauge the functionality of other crystallized
proteins with the molecular resolution of vibrational
spectroscopy.},
keywords = {Bacteriorhodopsins: chemistry / Bacteriorhodopsins:
radiation effects / Bacteriorhodopsins: ultrastructure /
Crystallography: methods / Dose-Response Relationship,
Radiation / Kinetics / Light / Photobiology: methods /
Photochemistry: methods / Radiation Dosage / Spectroscopy,
Fourier Transform Infrared: methods / Time Factors /
Bacteriorhodopsins (NLM Chemicals) / J (WoSType)},
cin = {IBI-2},
ddc = {570},
cid = {I:(DE-Juel1)VDB58},
pnm = {Funktion und Dysfunktion des Nervensystems},
pid = {G:(DE-Juel1)FUEK409},
shelfmark = {Biophysics},
typ = {PUB:(DE-HGF)16},
pubmed = {pmid:16731567},
pmc = {pmc:PMC1518640},
UT = {WOS:000239242000029},
doi = {10.1529/biophysj.106.083345},
url = {https://juser.fz-juelich.de/record/52270},
}