title: Monitoring Mitochondrial Pyruvate Carrier Activity in Real Time Using a BRET-Based Biosensor: Investigation of the Warburg Effect.
abstract: The transport of pyruvate into mitochondria requires a specific carrier, the mitochondrial pyruvate carrier (MPC). The MPC represents a central node of carbon metabolism, and its activity is likely to play a key role in bioenergetics. Until now, investigation of the MPC activity has been limited. However, the recent molecular identification of the components of the carrier has allowed us to engineer a genetically encoded biosensor and to monitor the activity of the MPC in real time in a cell population or in a single cell. We report that the MPC activity is low in cancer cells, which mainly rely on glycolysis to generate ATP, a characteristic known as the Warburg effect. We show that this low activity can be reversed by increasing the concentration of cytosolic pyruvate, thus increasing oxidative phosphorylation. This biosensor represents a unique tool to investigate carbon metabolism and bioenergetics in various cell types.
authors.affiliation: Department of Cell Biology, University of Geneva, 30 quai Ernest-Ansermet, 1211 Genève 4, Switzerland; Institut de Génomique Fonctionnelle Labex ICST; INSERM U661, CNRS UMR5203; Université Montpellier, 141, rue de la Cardonille, 34094 Montpellier cedex 5, France.
authors.forename: Vincent
authors.initials: V
authors.lastname: Compan
authors.affiliation: Department of Cell Biology, University of Geneva, 30 quai Ernest-Ansermet, 1211 Genève 4, Switzerland.
authors.forename: Sandra
authors.initials: S
authors.lastname: Pierredon
authors.affiliation: Department of Cell Biology, University of Geneva, 30 quai Ernest-Ansermet, 1211 Genève 4, Switzerland.
authors.forename: Benoît
authors.initials: B
authors.lastname: Vanderperre
authors.affiliation: ETH Zurich, Institute of Molecular Systems Biology Auguste-Piccard-Hof 1, 8093 Zurich, Switzerland.
authors.forename: Petra
authors.initials: P
authors.lastname: Krznar
authors.affiliation: Institute for Research on Cancer and Aging of Nice (IRCAN), University of Nice Sophia Antipolis, Centre A. Lacassagne, 06189 Nice, France.
authors.forename: Ibtissam
authors.initials: I
authors.lastname: Marchiq
authors.affiliation: ETH Zurich, Institute of Molecular Systems Biology Auguste-Piccard-Hof 1, 8093 Zurich, Switzerland.
authors.forename: Nicola
authors.initials: N
authors.lastname: Zamboni
authors.affiliation: Institute for Research on Cancer and Aging of Nice (IRCAN), University of Nice Sophia Antipolis, Centre A. Lacassagne, 06189 Nice, France; Centre Scientifique de Monaco, 8, Quai Antoine 1er MC, 98000, Monaco.
authors.forename: Jacques
authors.initials: J
authors.lastname: Pouyssegur
authors.affiliation: Department of Cell Biology, University of Geneva, 30 quai Ernest-Ansermet, 1211 Genève 4, Switzerland. Electronic address: jean-claude.martinou@unige.ch.
authors.forename: Jean-Claude
authors.initials: JC
authors.lastname: Martinou
mesh_terms: D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D002460:Cell Line; D004622:Embryo, Mammalian / Q000166:cytology; D004735:Energy Transfer; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism; D045325:HCT116 Cells; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D008163:Luminescent Measurements / Q000379:methods*; D061986:MCF-7 Cells; D051379:Mice; D033681:Mitochondrial Membrane Transport Proteins / Q000378:metabolism*; D019289:Pyruvic Acid / Q000378:metabolism*; D059010:Single-Cell Analysis
chemical_list: D033681:Mitochondrial Membrane Transport Proteins; D019289:Pyruvic Acid