Unveiling the K(+)-sensitivity of cell metabolism using genetically encoded, FRET-based K(+), glucose, and ATP biosensors

Investigating dynamic changes of mitochondrial ATP and cytosolic glucose levels of single living cells over time by genetically encoded biosensors provides an informative readout of their metabolic activities. Here, we describe how to monitor the metabolic K(+)-sensitivity of HEK293 cells exploiting...

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Detalles Bibliográficos
Autores principales: Bischof, Helmut, Burgstaller, Sandra, Graier, Wolfgang F., Lukowski, Robert, Malli, Roland
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2021
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8458979/
https://www.ncbi.nlm.nih.gov/pubmed/34589717
http://dx.doi.org/10.1016/j.xpro.2021.100843
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author Bischof, Helmut
Burgstaller, Sandra
Graier, Wolfgang F.
Lukowski, Robert
Malli, Roland
author_facet Bischof, Helmut
Burgstaller, Sandra
Graier, Wolfgang F.
Lukowski, Robert
Malli, Roland
author_sort Bischof, Helmut
collection PubMed
description Investigating dynamic changes of mitochondrial ATP and cytosolic glucose levels of single living cells over time by genetically encoded biosensors provides an informative readout of their metabolic activities. Here, we describe how to monitor the metabolic K(+)-sensitivity of HEK293 cells exploiting ATP-, glucose-, and K(+) probes. Fluorescence live-cell imaging of these Förster resonance energy transfer-based biosensors over time in response to gramicidin, an ionophoric peptide, indicated an absolute dependency of cellular ATP homeostasis on high intracellular K(+) levels. For complete information on the generation and use of this protocol please refer to Bischof et al. (2021).
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spelling pubmed-84589792021-09-28 Unveiling the K(+)-sensitivity of cell metabolism using genetically encoded, FRET-based K(+), glucose, and ATP biosensors Bischof, Helmut Burgstaller, Sandra Graier, Wolfgang F. Lukowski, Robert Malli, Roland STAR Protoc Protocol Investigating dynamic changes of mitochondrial ATP and cytosolic glucose levels of single living cells over time by genetically encoded biosensors provides an informative readout of their metabolic activities. Here, we describe how to monitor the metabolic K(+)-sensitivity of HEK293 cells exploiting ATP-, glucose-, and K(+) probes. Fluorescence live-cell imaging of these Förster resonance energy transfer-based biosensors over time in response to gramicidin, an ionophoric peptide, indicated an absolute dependency of cellular ATP homeostasis on high intracellular K(+) levels. For complete information on the generation and use of this protocol please refer to Bischof et al. (2021). Elsevier 2021-09-20 /pmc/articles/PMC8458979/ /pubmed/34589717 http://dx.doi.org/10.1016/j.xpro.2021.100843 Text en © 2021 The Authors https://creativecommons.org/licenses/by/4.0/This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Protocol
Bischof, Helmut
Burgstaller, Sandra
Graier, Wolfgang F.
Lukowski, Robert
Malli, Roland
Unveiling the K(+)-sensitivity of cell metabolism using genetically encoded, FRET-based K(+), glucose, and ATP biosensors
title Unveiling the K(+)-sensitivity of cell metabolism using genetically encoded, FRET-based K(+), glucose, and ATP biosensors
title_full Unveiling the K(+)-sensitivity of cell metabolism using genetically encoded, FRET-based K(+), glucose, and ATP biosensors
title_fullStr Unveiling the K(+)-sensitivity of cell metabolism using genetically encoded, FRET-based K(+), glucose, and ATP biosensors
title_full_unstemmed Unveiling the K(+)-sensitivity of cell metabolism using genetically encoded, FRET-based K(+), glucose, and ATP biosensors
title_short Unveiling the K(+)-sensitivity of cell metabolism using genetically encoded, FRET-based K(+), glucose, and ATP biosensors
title_sort unveiling the k(+)-sensitivity of cell metabolism using genetically encoded, fret-based k(+), glucose, and atp biosensors
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8458979/
https://www.ncbi.nlm.nih.gov/pubmed/34589717
http://dx.doi.org/10.1016/j.xpro.2021.100843
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