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

Andrei I. Tarasov; Guy A. Rutter

doi:10.1016/B978-0-12-416618-9.00015-7

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

Andrei I. Tarasov, Guy A. Rutter

Imperial College London

Research output: Chapter in Book/Report/Conference proceeding › Chapter › peer-review

24 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 Ca^{2 +} 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 Ca^{2 +} concentrations in malignant cells, two important aspects of oncometabolism.

Original language	English
Title of host publication	Conceptual Background and Bioenergetic/Mitochondrial Aspects of Oncometabolism
Publisher	Academic Press
Pages	289-311
Number of pages	23
ISBN (Print)	9780124166189
DOIs	https://doi.org/10.1016/B978-0-12-416618-9.00015-7
Publication status	Published (in print/issue) - 2014

Publication series

Name	Methods in Enzymology
Volume	542
ISSN (Print)	0076-6879
ISSN (Electronic)	1557-7988

Funding

A. I. T. holds an OxBRC fellowship. G. A. R. is supported by a Wellcome Trust Senior Investigator Award (WT098424AIA), MRC Programme (MR/J0003042/1), BBSRC (BB/J015873/1), and Diabetes UK (11/0004210) Project Grants and a Royal Society Wolfson Research Merit Award. The work leading to this publication has received support from the Innovative Medicines Initiative Joint Undertaking under grant agreement no. 155005 (IMIDIA), resources of which are composed of financial contribution from the European Union's Seventh Framework Programme (FP7/2007–2013) and EFPIA companies’ in-kind contribution (G. A. R.). We thank Dr. Gary Yellen (Harvard University) and members of the Rutter laboratory for useful discussion.

Keywords

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

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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10.1016/B978-0-12-416618-9.00015-7

Cite this

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KW - Glycolysis

KW - Live-cell imaging

KW - Pancreatic β-cells

KW - Patch clamp

KW - Wide-field microscopy

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ER -

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

Abstract

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Funding

Keywords

UN SDGs

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