Monitoring Mitochondrial Pyruvate Carrier Activity in Real Time Using a BRET-Based Biosensor: Investigation of the Warburg Effect

Mol Cell. 2015 Aug 6;59(3):491-501. doi: 10.1016/j.molcel.2015.06.035.

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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Biosensing Techniques / methods*
  • Cell Line
  • Embryo, Mammalian / cytology
  • Energy Transfer
  • Fibroblasts / cytology*
  • Fibroblasts / metabolism
  • HCT116 Cells
  • HEK293 Cells
  • HeLa Cells
  • Humans
  • Luminescent Measurements / methods*
  • MCF-7 Cells
  • Mice
  • Mitochondrial Membrane Transport Proteins / metabolism*
  • Pyruvic Acid / metabolism*
  • Single-Cell Analysis

Substances

  • Mitochondrial Membrane Transport Proteins
  • Pyruvic Acid