Evaluating the efficacy of GLUT inhibitors using a seahorse extracellular flux analyzer

Changyong Wei, Monique Heitmeier, Paul W. Hruz, Mala Shanmugam

Research output: Chapter in Book/Report/Conference proceedingChapter

4 Scopus citations

Abstract

Glucose is metabolized through anaerobic glycolysis and aerobic oxidative phosphorylation (OXPHOS). Perturbing glucose uptake and its subsequent metabolism can alter both glycolytic and OXPHOS pathways and consequently lactate and/or oxygen consumption. Production and secretion of lactate, as a consequence of glycolysis, leads to acidification of the extracellular medium. Molecular oxygen is the final electron acceptor in the electron transport chain, facilitating oxidative phosphorylation of ADP to ATP. The alterations in extracellular acidification and/or oxygen consumption can thus be used as indirect readouts of glucose metabolism and assessing the impact of inhibiting glucose transport through specific glucose transporters (GLUTs). The Seahorse bioenergetics analyzer can measure both the oxygen consumption rate (OCR) and extracellular acidification rate (ECAR). The proposed methodology affords a robust, high-throughput method to screen for GLUT inhibition in cells engineered to express specific GLUTs, providing live cell read-outs upon GLUT inhibition.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages69-75
Number of pages7
DOIs
StatePublished - Jan 1 2018

Publication series

NameMethods in Molecular Biology
Volume1713
ISSN (Print)1064-3745

Keywords

  • ECAR
  • GLUT inhibitor
  • Glycolysis
  • Oxidative phosphorylation
  • Seahorse XF analyzer

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    Wei, C., Heitmeier, M., Hruz, P. W., & Shanmugam, M. (2018). Evaluating the efficacy of GLUT inhibitors using a seahorse extracellular flux analyzer. In Methods in Molecular Biology (pp. 69-75). (Methods in Molecular Biology; Vol. 1713). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-7507-5_6