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Enhanced-acceptor fluorescence-based single cell ATP biosensor monitors ATP in heterogeneous cancer populations in real time

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Abstract

Current methods to monitor cellular ATP do not provide spatial or temporal localization of ATP in single cells in real time or they display imperfect specificity to ATP. Here, we have developed a single cell, Enhanced Acceptor Fluorescence (EAF)-based ATP biosensor to visualize ATP in real time. This biosensor utilizes a modified mimic of the ε-subunits of the Bacillus subtilis F0F1 synthase and is coupled to the EAF fluorophores pairs, GFP and YFP. The sensor was then used to monitor ATP in a heterogeneous glioblastoma multiform cancer cell population. We anticipate that this innovative technology and our better understanding of the ATP machinery will have substantial influence on future translational studies.

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References

  • Berg J, Hung YP, Yellen G (2009) A genetically encoded fluorescent reporter of ATP/ADP ratio. Nat Methods 2:161–166

    Article  Google Scholar 

  • Calleja V, Ameer-Beg SM, Vojnovic B, Woscholski R, Downwards J, Larijani B (2003) Monitoring conformational changes of proteins in cells by fluorescence’s lifetime imaging microscopy. Biochem J 372:33–40

    Article  PubMed  CAS  Google Scholar 

  • Das AM (2003) Regulation of the mitochondrial ATP-synthase in health and disease. Mol Genet Metab 2:71–82

    Article  Google Scholar 

  • DeAngelis LM (2001) Brain tumors. N Engl J Med 344:114–123

    Article  PubMed  CAS  Google Scholar 

  • Fang M, Chen Z, Huang S, Zhao L, Chen S, Mak TW, Wang X (2010) The ER UDPase ENTPD5 promotes protein N-glycosylation, the Warburg effect, and proliferation in the PTEN pathway. Cell 14:711–724

    Article  Google Scholar 

  • Glaysher S, Cree IA (2011) Cell sensitivity assays: the ATP-based tumor chemosensitivity assay. Methods Mol Bio 731:247–257

    Article  CAS  Google Scholar 

  • Hide I, Tanaka M, Inoue A, Nakajima K, Kohsaka S, Inoue K, Nakata Y (2000) Extracellular ATP triggers tumor necrosis factor-alpha release from rat microglia. J Neurochem 3:965–972

    Google Scholar 

  • Imamura H, Nhat KPH, Togawa H, Saito K, Lino R, Yamada YK, Nagai T, Noji H (2009) Visualization of ATP levels inside single living cells with fluorescent resonance energy transfer-based genetically encoded indicators. Proc Natl Acad Sci USA 37:15651–15656

    Article  Google Scholar 

  • Israelsen WJ, Vander Heiden MG (2010) ATP consumption promotes cancer metabolism. Cell 143:669–671

    Article  PubMed  CAS  Google Scholar 

  • Locasale JW, Cantley LC (2012) Altered metabolism in cancer. BMC Biol 8:88

    Article  Google Scholar 

  • Parsons DW, Jones S, Zhang X, Lin JC, Leary RJ, Angenendt P (2008) An integrated genomic analysis of human glioblastoma multiform. Science 5897:1807–1812

    Article  Google Scholar 

  • Pérez-Flores I, Sánchez-Fructuoso A, Santiago JL, Fernández-Arquero M, Calvo N, Gómez de la Concha E, Barrientos A (2009) Intracellular ATP levels in CD4+ lymphocytes are a risk marker of rejection and infection in renal graft recipients. Transpl Proc 6:2106–2108

    Article  Google Scholar 

  • Stephenson K, Hoch JA (2001) PAS-A domain of phosphorelay sensor kinase A: a catalytic ATP-binding domain involved in the initiaton of development in Bacillus subtilis. Proc Natl Acad Sci USA 98:15251–15256

    Article  PubMed  CAS  Google Scholar 

  • Tsujimoto Y (1997) Apoptosis and necrosis: intracellular ATP as a determinant for cell death modes. Cell Death Differ 6:429–434

    Article  Google Scholar 

  • Vander Heiden MG, Cantley L, Thompson C (2009) Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science 5930:1029–1033

    Article  Google Scholar 

  • Zadran S, Qin Q, Bi X, Zadran H, Kim Y, Foy MR, Thompson R, Baudry M (2009) 17-B Estradiol increases neuronal excitability through MAP kinase-induced calpain activation. Proc Natl Acad Sci USA 51:21936–21941

    Article  Google Scholar 

Download references

Acknowledgments

We would also like to thank Dr. Raphael Levine (UCLA) and Dr. Michel Baudry (USC) for critical analysis and discussions. We would like to thank the UCLA Brain Tumor Translational Resource for biospecimen support. The UCLA David Geffen School of Medicine for early career funds to S. Zadran and the National Science Foundation Graduate Research Fellowship Program (GRFP) to H. Zadran supported this work.

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Authors and Affiliations

  1. David Geffen School of Medicine, University of California, Los Angeles, 10833 Le Conte Ave., CHS 33-165, Los Angeles, CA, 90089, USA

    Sohila Zadran, Desiree Sanchez, Arash Amighi, Erick Otiniano & Kaylee Wong

  2. Department of Biological Sciences, University of Southern California, Los Angeles, 3461 Watt Way, HNB 319, Los Angeles, CA, 90089, USA

    Homera Zadran

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  1. Sohila Zadran
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  2. Desiree Sanchez
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  3. Homera Zadran
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  4. Arash Amighi
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  5. Erick Otiniano
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  6. Kaylee Wong
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Correspondence to Sohila Zadran.

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Zadran, S., Sanchez, D., Zadran, H. et al. Enhanced-acceptor fluorescence-based single cell ATP biosensor monitors ATP in heterogeneous cancer populations in real time. Biotechnol Lett 35, 175–180 (2013). https://doi.org/10.1007/s10529-012-1065-6

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  • DOI: https://doi.org/10.1007/s10529-012-1065-6

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