Substrate Binding Affinity Changes in Mitochondrial Energy-linked Reactions

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1982 Mar 1

PMID 6952227

Citations 6

Authors

Y HATEFI

T Yagi

D C Phelps

S Y Wong

S B Vik

Y M Galante

Affiliations

Soon will be listed here.

Abstract

The effects of uncouplers and valinomycin plus nigericin (in the presence of K+) were studied on the apparent Km for substrates and apparent Vmax of the following energy-linked reactions catalyzed by submitochondrial particles: oxidative phosphorylation, NTP-33Pi exchange, ATP-driven electron transfer from succinate to NAD, and respiration-driven transhydrogenation from NADH to 3-acetylpyridine adenine dinucleotide phosphate. In all cases, partially uncoupling (up to 90%) concentrations of uncouplers of valinomycin plus nigericin were found to decrease apparent Vmax and to increase apparent Km. Results plotted as ln (Vmax/Km) versus the concentration of uncouplers or ionophores showed a linear decrease of the former as a function of increasing perturbant concentration (i.e., decreasing free energy). Because Vmax/Km may be considered as a measure of the apparent first-order rate constant for enzyme-substrate interaction and reflects the affinity between enzyme and substrate to form a complex, the results are consistent with the interpretation that membrane energization leads to a change in enzyme conformation with the resultant increase in enzyme-substrate affinity and facilitation of the reaction rate under consideration. The significance of these findings with respect to the mechanism of action of the energy-transducing systems studied is discussed.

Citing Articles

Steady-state kinetics of the overall oxidative phosphorylation reaction in heart mitochondria. Determination of the coupling relationships between the respiratory reactions and miscellaneous observations concerning rate-limiting steps.

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A critical appraisal of evidence for localized energy coupling. Kinetic studies on liposomes containing bacteriorhodopsin and ATP synthase.

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Role of energy in oxidative phosphorylation.

HATEFI Y J Bioenerg Biomembr. 1988; 20(4):481-502.

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Fluorometric evidence for control of the activity of F1-adenosinetriphosphatase by ligand-induced conformation change.

Wang J J Bioenerg Biomembr. 1986; 18(2):101-11.

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Studies on the mechanism of oxidative phosphorylation: effects of specific F0 modifiers on ligand-induced conformation changes of F1.

Yagi T, HATEFI Y Proc Natl Acad Sci U S A. 1985; 82(22):7550-4.

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