A genetically encoded single-wavelength sensor for imaging cytosolic and cell surface ATP.
basic_science · Level V
Where this comes from
- Record sourced from PubMed, PMID 30755613.
- Also identified by DOI 10.1038/s41467-019-08441-5 and PMC identifier 6372613.
- Licence recorded as CC BY.
- The licence permits redistribution, so the abstract is shown in full and the full text is available from the publisher.
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 F<sub>0</sub>F<sub>1</sub>-ATPase from Bacillus PS3. On the cell surface and within the cytosol, iATPSnFR<sup>1.0</sup> 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.
Medical subject headings
- Adenosine Triphosphate
- Cell Membrane
- Cytosol
- Fluorescence Resonance Energy Transfer
- Proteins