Nature communications, 2019-02, Vol.10 (1), p.711-13, Article 711
2019
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Details
- Autor(en) / Beteiligte
- Titel
- A genetically encoded single-wavelength sensor for imaging cytosolic and cell surface ATP
- Ist Teil von
- Nature communications, 2019-02, Vol.10 (1), p.711-13, Article 711
- Ort / Verlag
- England: Nature Publishing Group
- Erscheinungsjahr
- 2019
- Quelle
- Free E-Journal (出版社公開部分のみ）
- Beschreibungen/Notizen
- 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 F -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.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 2041-1723eISSN: 2041-1723DOI: 10.1038/s41467-019-08441-5Titel-ID: cdi_doaj_primary_oai_doaj_org_article_7f5127aea1404556aa178bc1605413dd
- Format
- –
- Schlagworte
- Adenosine, Adenosine triphosphatase, Adenosine triphosphate, Adenosine Triphosphate - chemistry, Adenosine Triphosphate - genetics, Amino acids, ATP, ATPase Inhibitory Protein, Bacillus - cytology, Bacillus - genetics, Bacillus - metabolism, Bacterial Proteins - genetics, Cell Membrane - chemistry, Cell surface, Cloning, Coding, Cytosol, Cytosol - chemistry, F0F1-ATPase, Fluorescence, Fluorescence Resonance Energy Transfer - methods, Gene Expression, Genetic code, Green fluorescent protein, HEK293 Cells, Homeostasis, Humans, Hydrogen-Ion Concentration, Image Processing, Computer-Assisted, Imaging, Insertion, Kinetics, Luminescent Proteins - genetics, Microscopes, Microscopy, Fluorescence, Models, Molecular, Mutagenesis, Site-Directed, Neurosciences, Nucleotides, Protein Conformation, Proteins, Proteins - metabolism, Reagents, Red Fluorescent Protein, Sensors, Wavelength