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

Lobas, M.; Nagai, J.; Kronschlager, M. T.; Borden, P.; Marvin, J. S.; Looger, L. L.; Khakh, B. S.

doi:10.1101/385484

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bioRxiv · 10.1101/385484

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

Lobas, M.·Nagai, J.·Kronschlager, M. T.·Borden, P.·Marvin, J. S.·Looger, L. L.·Khakh, B. S.

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Abstract

Adenosine 5 triphosphate (ATP) is a universal intracellular energy source1 and an evolutionarily ancient2 extracellular signal3-5. 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 responded to relevant ATP concentrations (30 M 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. iATPSnFRs represent promising new reagents for imaging ATP dynamics.

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Lobas, M., Nagai, J., Kronschlager, M. T., Borden, P., Marvin, J. S., Looger, L. L., Khakh, B. S.. 2018-08-06. A genetically encoded single-wavelength sensor for imaging cytosolic and cell surface ATP. https://doi.org/10.1101/385484

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