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Direct measurement of energy fluxes from mitochondria into cytoplasm in permeabilized cardiac cells in situ: some evidence for mitochondrial interactosome

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Abstract

The aim of this study was to measure energy fluxes from mitochondria in isolated permeabilized cardiomyocytes. Respiration of permeabilized cardiomyocytes and mitochondrial membrane potential were measured in presence of MgATP, pyruvate kinase – phosphoenolpyruvate and creatine. ATP and phosphocreatine concentrations in medium surrounding cardiomyocytes were determined. While ATP concentration did not change in time, mitochondria effectively produced phosphocreatine (PCr) with PCr/O2 ratio equal to 5.68 ± 0.14. Addition of heterodimeric tubulin to isolated mitochondria was found to increase apparent Km for exogenous ADP from 11 ± 2 µM to 330 ± 47 µM, but creatine again decreased it to 23 ± 6 µM. These results show directly that under physiological conditions the major energy carrier from mitochondria into cytoplasm is PCr, produced by mitochondrial creatine kinase (MtCK), which functional coupling to adenine nucleotide translocase is enhanced by selective limitation of permeability of mitochondrial outer membrane within supercomplex ATP Synthasome-MtCK-VDAC-tubulin, Mitochondrial Interactosome.

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  1. Laboratory of Bioenergetics, National Institute of Chemical Physics and Biophysics, Tallinn, Estonia

    Natalia Timohhina, Kersti Tepp, Minna Varikmaa, Heiki Vija, Peeter Sikk, Tuuli Kaambre & Valdur Saks

  2. Laboratory of Fundamental and Applied Bioenergetics, INSERM U884, Joseph Fourier University, 2280, Rue de la Piscine, BP53X – 38041, Grenoble Cedex 9, France

    Rita Guzun, Claire Monge & Valdur Saks

  3. Laboratory of Integrative and Medical Biophysics, National Institute of Child Health and Human Development, National Institutes of Health, Bethesda, MD, USA

    Dan Sackett

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  1. Natalia Timohhina
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Timohhina, N., Guzun, R., Tepp, K. et al. Direct measurement of energy fluxes from mitochondria into cytoplasm in permeabilized cardiac cells in situ: some evidence for mitochondrial interactosome. J Bioenerg Biomembr 41, 259–275 (2009). https://doi.org/10.1007/s10863-009-9224-8

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