Regulation of glycolytic oscillations by mitochondrial and plasma membrane H+-ATPases

Lars Folke Olsen, Ann Zahle Andersen, Anita Lunding, Jens Christian Brasen, Allan K Poulsen

Research output: Contribution to journalJournal articleResearchpeer-review

Abstract

We investigated the coupling between glycolytic and mitochondrial membrane potential oscillations in Saccharomyces cerevisiae under semianaerobic conditions. Glycolysis was measured as NADH autofluorescence, and mitochondrial membrane potential was measured using the fluorescent dye 3,3'-diethyloxacarbocyanine iodide. The responses of glycolytic and membrane potential oscillations to a number of inhibitors of glycolysis, mitochondrial electron flow, and mitochondrial and plasma membrane H(+)-ATPase were investigated. Furthermore, the glycolytic flux was determined as the rate of production of ethanol in a number of different situations (changing pH or the presence and absence of inhibitors). Finally, the intracellular pH was determined and shown to oscillate. The results support earlier work suggesting that the coupling between glycolysis and mitochondrial membrane potential is mediated by the ADP/ATP antiporter and the mitochondrial F(0)F(1)-ATPase. The results further suggest that ATP hydrolysis, through the action of the mitochondrial F(0)F(1)-ATPase and plasma membrane H(+)-ATPase, are important in regulating these oscillations. We conclude that it is glycolysis that drives the oscillations in mitochondrial membrane potential.
Original languageEnglish
JournalBIOPHYSICAL JOURNAL
Volume96
Issue number9
Pages (from-to)3850-61
ISSN0006-3495
DOIs
Publication statusPublished - 2009
Externally publishedYes

Keywords

  • Adenosine Triphosphate
  • Carbon Dioxide
  • Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone
  • Cell Membrane
  • Dithiazanine
  • Enzyme Inhibitors
  • Ethanol
  • Fluorescence
  • Glycolysis
  • Hydrogen-Ion Concentration
  • Membrane Potential, Mitochondrial
  • Membrane Potentials
  • Mitochondrial Membranes
  • NAD
  • Omeprazole
  • Oxygen Consumption
  • Periodicity
  • Proton-Translocating ATPases
  • Saccharomyces cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Uncoupling Agents

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