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J. Biol. Chem., Vol. 263, Issue 6, 2712-2718, Feb, 1988
DF Wilson, WL Rumsey, TJ Green and JM Vanderkooi
Oxygen-dependent quenching of phosphorescence has been used to measure the
dependence of mitochondrial oxidative phosphorylation on oxygen
concentration in suspensions of isolated rat liver mitochondria. An
instrument has been designed which simultaneously monitors the
phosphorescence lifetime of a fluorophor and the reduction of cytochrome c
by dual wavelength spectrophotometry. The phosphorescence lifetime method
gives very rapid (less than 100 ms) measure of the oxygen concentration
(Vanderkooi, J. M., Maniara, G., Green, T. J., and Wilson, D. F. (1987) J.
Biol. Chem. 262, 5476-5482) from concentrations characteristic of
air-saturated media to as low as 2 x 10(-8) M. The results may be
summarized as follows. For well coupled rat liver mitochondria at pH 7.0
and in the presence of ATP, as the oxygen concentration was lowered,
increased cytochrome c reduction was observed to begin at oxygen
concentrations greater than 20 microM. For mitochondria in the presence of
uncoupler, cytochrome c reduction began at oxygen concentrations less than
1.0 microM. The oxygen dependence of reduction of cytochrome c in well
coupled mitochondria treated with ATP was strongly dependent on the pH of
the suspending medium. Reduction of cytochrome c began at higher oxygen
concentrations as the pH was made more alkaline. The oxygen concentration
for half-maximal respiratory rates was much larger for well coupled
mitochondria treated with ATP (approximately 0.7 microM) than for
mitochondria treated with uncoupler (less than 0.1 microM). It is concluded
that the oxygen dependence of mitochondrial oxidative phosphorylation is
such that mitochondria could function in their proposed role of tissue
oxygen sensors for regulation of such diverse functions as local blood flow
and electrical activity in the carotid body.
The oxygen dependence of mitochondrial oxidative phosphorylation measured by a new optical method for measuring oxygen concentration
Department of Biochemistry and Biophysics, Medical School, University of Pennsylvania, Philadelphia 19104.
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