Use of genetically encoded sensors to monitor cytosolic ATP/ADP ratio in living cells

Andrei I. Tarasov, Guy A. Rutter

Research output: Chapter in Book/Report/Conference proceedingChapter

9 Citations (Scopus)

Abstract

ATP is not only recognized as the universal energy "currency" in most cells but also plays a less well-known role as an intracellular and extracellular messenger. Here, we review novel approaches for measuring free ATP (or ATP/ADP ratios) in living mammalian cells by using genetically encoded sensors. We also discuss the key technical aspects of routine real-time ATP/ADP monitoring using as a model one of the last-generation fluorescent probes, a fusion protein commonly known as "Perceval." Finally, we present detailed guidelines for the simultaneous measurement of cytosolic ATP/ADP ratios and Ca2 + concentrations alongside electrical parameters in individual pancreatic β cells, in which energy metabolism is tightly linked to plasma membrane excitability to control the secretion of insulin. With appropriate variations, this approach can be adapted to the study of cytosolic ATP/ADP ratios and Ca2 + concentrations in malignant cells, two important aspects of oncometabolism.

Original languageEnglish
Title of host publicationConceptual Background and Bioenergetic/Mitochondrial Aspects of Oncometabolism
PublisherAcademic Press
Pages289-311
Number of pages23
ISBN (Print)9780124166189
DOIs
Publication statusPublished - 2014

Publication series

NameMethods in Enzymology
Volume542
ISSN (Print)0076-6879
ISSN (Electronic)1557-7988

Keywords

  • Cytosolic ATP
  • Glycolysis
  • Live-cell imaging
  • Pancreatic β-cells
  • Patch clamp
  • Wide-field microscopy

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    Tarasov, A. I., & Rutter, G. A. (2014). Use of genetically encoded sensors to monitor cytosolic ATP/ADP ratio in living cells. In Conceptual Background and Bioenergetic/Mitochondrial Aspects of Oncometabolism (pp. 289-311). (Methods in Enzymology; Vol. 542). Academic Press. https://doi.org/10.1016/B978-0-12-416618-9.00015-7