PT  - JOURNAL ARTICLE
AU  - Mark Lobas
AU  - Jun Nagai
AU  - Mira T. Kronschläger
AU  - Philip Borden
AU  - Jonathan S. Marvin
AU  - Loren L. Looger
AU  - Baljit S. Khakh
TI  - A genetically encoded single-wavelength sensor for imaging cytosolic and cell surface ATP
AID  - 10.1101/385484
DP  - 2018 Jan 01
TA  - bioRxiv
PG  - 385484
4099  - http://biorxiv.org/content/early/2018/08/06/385484.short
4100  - http://biorxiv.org/content/early/2018/08/06/385484.full
AB  - 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 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. iATPSnFRs represent promising new reagents for imaging ATP dynamics.