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

Mark A. Lobas, Rongkun Tao, Jun Nagai, Mira T. Kronschläger, Philip M. Borden, Jonathan S. Marvin, Loren L. Looger () and Baljit S. Khakh ()
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Mark A. Lobas: University of California Los Angeles
Rongkun Tao: University of California Los Angeles
Jun Nagai: University of California Los Angeles
Mira T. Kronschläger: University of California Los Angeles
Philip M. Borden: Janelia Research Campus
Jonathan S. Marvin: Janelia Research Campus
Loren L. Looger: Janelia Research Campus
Baljit S. Khakh: University of California Los Angeles

Nature Communications, 2019, vol. 10, issue 1, 1-13

Abstract: Abstract 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 F0F1-ATPase from Bacillus PS3. On the cell surface and within the cytosol, iATPSnFR1.0 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.

Date: 2019
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DOI: 10.1038/s41467-019-08441-5

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