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A genetically encoded single-wavelength sensor for imaging cytosolic and cell surface ATP

Adenosine 5′ triphosphate (ATP) is a universal intracellular energy source and an evolutionarily ancient, ubiquitous extracellular signal in diverse species. Here, we report the generation and characterization of single-wavelength genetically encoded fluorescent sensors (iATPSnFRs) for imaging extra...

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Autores principales: Lobas, Mark A., Tao, Rongkun, Nagai, Jun, Kronschläger, Mira T., Borden, Philip M., Marvin, Jonathan S., Looger, Loren L., Khakh, Baljit S.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6372613/
https://www.ncbi.nlm.nih.gov/pubmed/30755613
http://dx.doi.org/10.1038/s41467-019-08441-5
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author Lobas, Mark A.
Tao, Rongkun
Nagai, Jun
Kronschläger, Mira T.
Borden, Philip M.
Marvin, Jonathan S.
Looger, Loren L.
Khakh, Baljit S.
author_facet Lobas, Mark A.
Tao, Rongkun
Nagai, Jun
Kronschläger, Mira T.
Borden, Philip M.
Marvin, Jonathan S.
Looger, Loren L.
Khakh, Baljit S.
author_sort Lobas, Mark A.
collection PubMed
description Adenosine 5′ triphosphate (ATP) is a universal intracellular energy source and an evolutionarily ancient, ubiquitous extracellular signal in diverse species. Here, we report the generation and characterization of single-wavelength genetically encoded fluorescent sensors (iATPSnFRs) for imaging extracellular and cytosolic ATP from insertion of circularly permuted superfolder GFP into the epsilon subunit of F(0)F(1)-ATPase from Bacillus PS3. On the cell surface and within the cytosol, iATPSnFR(1.0) responds to relevant ATP concentrations (30 μM to 3 mM) with fast increases in fluorescence. iATPSnFRs can be genetically targeted to specific cell types and sub-cellular compartments, imaged with standard light microscopes, do not respond to other nucleotides and nucleosides, and when fused with a red fluorescent protein function as ratiometric indicators. After careful consideration of their modest pH sensitivity, iATPSnFRs represent promising reagents for imaging ATP in the extracellular space and within cells during a variety of settings, and for further application-specific refinements.
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spelling pubmed-63726132019-02-14 A genetically encoded single-wavelength sensor for imaging cytosolic and cell surface ATP Lobas, Mark A. Tao, Rongkun Nagai, Jun Kronschläger, Mira T. Borden, Philip M. Marvin, Jonathan S. Looger, Loren L. Khakh, Baljit S. Nat Commun Article Adenosine 5′ triphosphate (ATP) is a universal intracellular energy source and an evolutionarily ancient, ubiquitous extracellular signal in diverse species. Here, we report the generation and characterization of single-wavelength genetically encoded fluorescent sensors (iATPSnFRs) for imaging extracellular and cytosolic ATP from insertion of circularly permuted superfolder GFP into the epsilon subunit of F(0)F(1)-ATPase from Bacillus PS3. On the cell surface and within the cytosol, iATPSnFR(1.0) responds to relevant ATP concentrations (30 μM to 3 mM) with fast increases in fluorescence. iATPSnFRs can be genetically targeted to specific cell types and sub-cellular compartments, imaged with standard light microscopes, do not respond to other nucleotides and nucleosides, and when fused with a red fluorescent protein function as ratiometric indicators. After careful consideration of their modest pH sensitivity, iATPSnFRs represent promising reagents for imaging ATP in the extracellular space and within cells during a variety of settings, and for further application-specific refinements. Nature Publishing Group UK 2019-02-12 /pmc/articles/PMC6372613/ /pubmed/30755613 http://dx.doi.org/10.1038/s41467-019-08441-5 Text en © The Author(s) 2019 Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
spellingShingle Article
Lobas, Mark A.
Tao, Rongkun
Nagai, Jun
Kronschläger, Mira T.
Borden, Philip M.
Marvin, Jonathan S.
Looger, Loren L.
Khakh, Baljit S.
A genetically encoded single-wavelength sensor for imaging cytosolic and cell surface ATP
title A genetically encoded single-wavelength sensor for imaging cytosolic and cell surface ATP
title_full A genetically encoded single-wavelength sensor for imaging cytosolic and cell surface ATP
title_fullStr A genetically encoded single-wavelength sensor for imaging cytosolic and cell surface ATP
title_full_unstemmed A genetically encoded single-wavelength sensor for imaging cytosolic and cell surface ATP
title_short A genetically encoded single-wavelength sensor for imaging cytosolic and cell surface ATP
title_sort genetically encoded single-wavelength sensor for imaging cytosolic and cell surface atp
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6372613/
https://www.ncbi.nlm.nih.gov/pubmed/30755613
http://dx.doi.org/10.1038/s41467-019-08441-5
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