Main Menu (Mobile)- Block

Main Menu - Block

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

Our Research / Publications
janelia7_blocks-janelia7_biblio_header | block
Nature Communications. 2019 Feb 12;10(1):711. doi: 10.1038/s41467-019-08441-5
View Article on
A genetically encoded single-wavelength sensor for imaging cytosolic and cell surface ATP. Looger Lab
Lobas MA, Tao R, Nagai J, Kronschläger MT, Borden PM, Marvin JS, Looger LL, Khakh BS
janelia7_blocks-janelia7_biblio_abstract | block

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 FF-ATPase from Bacillus PS3. On the cell surface and within the cytosol, iATPSnFR 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.

PMID: 30755613 [PubMed - indexed for MEDLINE]

node:body | entity_field
janelia7_blocks-janelia7_biblio_authors | block

Janelia Authors

Jonathan Marvin
Loren Looger
janelia7_blocks-janelia7_biblio_tools | block