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@ARTICLE{Heitbrink:58498,
author = {Heitbrink, D. and Sigurdson, H. and Bolwien, C. and
Brzezinski, P. and Heberle, J.},
title = {{T}ransient binding of {CO} to {C}u({B}) in cytochrome c
oxidase is dynamically linked to structural changes around a
carboxyl group : a time-resolved step-scan {F}ourier
transform infrared investigation},
journal = {Biophysical journal},
volume = {82},
issn = {0006-3495},
address = {New York, NY},
publisher = {Rockefeller Univ. Press},
reportid = {PreJuSER-58498},
pages = {1 - 10},
year = {2002},
note = {Record converted from VDB: 12.11.2012},
abstract = {The redox-driven proton pump cytochrome c oxidase is that
enzymatic machinery of the respiratory chain that transfers
electrons from cytochrome c to molecular oxygen and thereby
splits molecular oxygen to form water. To investigate the
reaction mechanism of cytochrome c oxidase on the single
vibrational level, we used time-resolved step-scan Fourier
transform infrared spectroscopy and studied the dynamics of
the reduced enzyme after photodissociation of bound carbon
monoxide across the midinfrared range (2300-950 cm(-1)).
Difference spectra of the bovine complex were obtained at
-20degreesC with 5 mus time resolution. The data demonstrate
a dynamic link between the transient binding of CO to Cu-B
and changes in hydrogen bonding at the functionally
important residue E(I-286). Variation of the pH revealed
that the pK(a) of E(I-286) is >9.3 in the fully reduced
CO-bound oxidase. Difference spectra of cytochrome c oxidase
from beef heart are compared with those of the oxidase
isolated from Rhodobacter sphaeroides. The bacterial enzyme
does not show the environmental change in the vicinity of
E(I-286) upon CO dissociation. The characteristic band shape
appears, however, in redox-induced difference spectra of the
bacterial enzyme but is absent in redox-induced difference
spectra of mammalian enzyme. In conclusion, it is
demonstrated that the dynamics of a large protein complex
such as cytochrome c oxidase can be resolved on the single
vibrational level with microsecond Fourier transform
infrared spectroscopy. The applied methodology provides the
basis for future investigations of the physiological
reaction steps of this important enzyme.},
keywords = {J (WoSType)},
cin = {IBI-2},
ddc = {570},
cid = {I:(DE-Juel1)VDB58},
pnm = {Neurowissenschaften},
pid = {G:(DE-Juel1)FUEK255},
shelfmark = {Biophysics},
typ = {PUB:(DE-HGF)16},
UT = {WOS:000173250500001},
url = {https://juser.fz-juelich.de/record/58498},
}