Regulation of glycolytic oscillations by mitochondrial and plasma membrane H+-ATPases.
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.
Olsen
Lars Folke
LF
CelCom, Department of Biochemistry and Molecular Biology, University of Southern Denmark, Odense, Denmark. lfo@bmb.sdu.dk
Andersen
Ann Zahle
AZ
Lunding
Anita
A
Brasen
Jens Christian
JC
Poulsen
Allan K
AK
D000255:Adenosine Triphosphate / Q000378:metabolism; D002245:Carbon Dioxide / Q000378:metabolism; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / Q000494:pharmacology; D002462:Cell Membrane / Q000502:physiology*; D004225:Dithiazanine; D004791:Enzyme Inhibitors / Q000494:pharmacology; D000431:Ethanol / Q000378:metabolism; D005453:Fluorescence; D006019:Glycolysis; D006863:Hydrogen-Ion Concentration; D053078:Membrane Potential, Mitochondrial / Q000502:physiology; D008564:Membrane Potentials / Q000502:physiology; D051336:Mitochondrial Membranes / Q000502:physiology*; D009243:NAD / Q000378:metabolism; D009853:Omeprazole / Q000494:pharmacology; D010101:Oxygen Consumption; D010507:Periodicity; D006180:Proton-Translocating ATPases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D012441:Saccharomyces cerevisiae; D029701:Saccharomyces cerevisiae Proteins / Q000378:metabolism; D014475:Uncoupling Agents / Q000494:pharmacology
D004791:Enzyme Inhibitors; D029701:Saccharomyces cerevisiae Proteins; D014475:Uncoupling Agents; D009243:NAD; D002245:Carbon Dioxide; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone; D000431:Ethanol; D004225:Dithiazanine; D000255:Adenosine Triphosphate; D006180:Proton-Translocating ATPases; D009853:Omeprazole