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

Author

Listed:
  • Mark A. Lobas

    (University of California Los Angeles
    Koniku Inc.)

  • Rongkun Tao

    (University of California Los Angeles)

  • Jun Nagai

    (University of California Los Angeles)

  • Mira T. Kronschläger

    (University of California Los Angeles
    Medical University of Vienna)

  • Philip M. Borden

    (Janelia Research Campus)

  • Jonathan S. Marvin

    (Janelia Research Campus)

  • Loren L. Looger

    (Janelia Research Campus)

  • Baljit S. Khakh

    (University of California Los Angeles
    University of California Los Angeles)

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 F0F1-ATPase from Bacillus PS3. On the cell surface and within the cytosol, iATPSnFR1.0 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.

Suggested Citation

  • Mark A. Lobas & Rongkun Tao & Jun Nagai & Mira T. Kronschläger & Philip M. Borden & Jonathan S. Marvin & Loren L. Looger & Baljit S. Khakh, 2019. "A genetically encoded single-wavelength sensor for imaging cytosolic and cell surface ATP," Nature Communications, Nature, vol. 10(1), pages 1-13, December.
    • Handle: RePEc:nat:natcom:v:10:y:2019:i:1:d:10.1038_s41467-019-08441-5
    DOI: 10.1038/s41467-019-08441-5
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    Cited by:

    1. Danielle L. Schmitt & Stephanie D. Curtis & Anne C. Lyons & Jin-fan Zhang & Mingyuan Chen & Catherine Y. He & Sohum Mehta & Reuben J. Shaw & Jin Zhang, 2022. "Spatial regulation of AMPK signaling revealed by a sensitive kinase activity reporter," Nature Communications, Nature, vol. 13(1), pages 1-12, December.
    2. Dorothy Koveal & Paul C. Rosen & Dylan J. Meyer & Carlos Manlio Díaz-García & Yongcheng Wang & Li-Heng Cai & Peter J. Chou & David A. Weitz & Gary Yellen, 2022. "A high-throughput multiparameter screen for accelerated development and optimization of soluble genetically encoded fluorescent biosensors," Nature Communications, Nature, vol. 13(1), pages 1-14, December.
    3. Shannon Trombley & Jackson Powell & Pavithran Guttipatti & Andrew Matamoros & Xiaohui Lin & Tristan O’Harrow & Tobias Steinschaden & Leann Miles & Qin Wang & Shuchao Wang & Jingyun Qiu & Qingyang Li &, 2023. "Glia instruct axon regeneration via a ternary modulation of neuronal calcium channels in Drosophila," Nature Communications, Nature, vol. 14(1), pages 1-18, December.

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