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High-throughput Live Cell Imaging and Analysis for Temporal Reaction of G Protein-coupled Receptor Based on Split Luciferase Fragment Complementation

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Abstract

For the design of therapeutic drugs, G protein coupled receptors (GPCRs) are notable targets. Many screening methods have been developed to identify effective agents for GPCR signaling. However, analyses of temporal variations of GPCR activity with specific ligands remain insufficient because of monitoring method limitations and difficulties. We previously developed a high-throughput bioluminescence measuring system to detect interactions of GPCR with β-arrestin based on split luciferase fragment complementation. By newly introducing a bioluminescence imaging technique into the system, we demonstrate a method for the temporal monitoring of GPCR–β-arrestin interactions in living cells during stimulation by different ligands.

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References

  1. R. Grisshammer, Expert Rev. Proteomics, 2013, 10, 1.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. P. J. Conn, A. Christopoulos, and C. W. Lindsley, Nat. Rev. Drug Discovery, 2009, 8, 41.

    Article  CAS  PubMed  Google Scholar 

  3. A. Musnier, B. Blanchot, E. Reiter, and P. Crepieux, Cell. Signalling, 2010, 22, 707.

    Article  CAS  PubMed  Google Scholar 

  4. R. C. Stevens, V. Cherezov, V. Katritch, R. Abagyan, P. Kuhn, H. Rosan, and K. Wüthrich, Nat. Rev. Drug Discovery, 2013, 12, 25.

    Article  CAS  PubMed  Google Scholar 

  5. A. L. Hopkins and C. R. Groom, Nat. Rev. Drug Discovery, 2002, 1, 727.

    Article  CAS  PubMed  Google Scholar 

  6. S. M. Foord, T. I. Bonner, R. R. Neubig, E. M. Rosser, J. P. Pin, A. P. Davenport, M. Spedding, and A. J. Harmar, Pharmacol. Rev., 2005, 57, 279.

    Article  CAS  PubMed  Google Scholar 

  7. R. Zhang and X. Xie, Acta Pharmacol. Sin., 2012, 33, 372.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. M. Hattori, M. Tanaka, H. Takakura, K. Aoki, K. Miura, T. Anzai, and T. Ozawa, Mol. BioSyst., 2013, 9, 957.

    Article  CAS  PubMed  Google Scholar 

  9. N. Hida, M. Awais, M. Takeuchi, N. Ueno, M. Tashiro, C. Takagi, T. Singh, M. Hayashi, Y. Ohmiya, and T. Ozawa, PLoS One, 2009, 4, e5868.

    Article  PubMed  PubMed Central  Google Scholar 

  10. N. Misawa, A. K. M. Kafi, M. Hattori, K. Miura, and T. Ozawa, Anal. Chem., 2010, 82, 2552.

    Article  CAS  PubMed  Google Scholar 

  11. H. Takakura, M. Hattori, M. Takeuchi, and T. Ozawa, ACS Chem. Biol., 2012, 7, 901.

    Article  CAS  PubMed  Google Scholar 

  12. A. K. M. Kafi, M. Hattori, N. Misawa, and T. Ozawa, Pharmaceuticals, 2011, 4, 457.

    Article  CAS  PubMed Central  Google Scholar 

  13. M. A. Paley and J. A. Prescher, Med. Chem. Commun., 2014, 5, 255.

    Article  CAS  Google Scholar 

  14. M. Yasunaga, Y. Nakajima, and Y. Ohmiya, Anal. Bioanal. Chem., 2014, 406, 5735.

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

We thank Molecular Devices Corp. for advice and support related to the technical side of imaging. This work was supported financially by the Japan Society for the Promotion of Science (JSPS), Japan Science and Technology (JST), and MEXT, Japan.

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Authors and Affiliations

  1. Department of Chemistry, School of Science, The University of Tokyo, 7-3-1 Hongo, Bunkyo, Tokyo, 113-0033, Japan

    Mitsuru Hattori & Takeaki Ozawa

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  1. Mitsuru Hattori
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  2. Takeaki Ozawa
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Correspondence to Mitsuru Hattori.

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Hattori, M., Ozawa, T. High-throughput Live Cell Imaging and Analysis for Temporal Reaction of G Protein-coupled Receptor Based on Split Luciferase Fragment Complementation. ANAL. SCI. 31, 327–330 (2015). https://doi.org/10.2116/analsci.31.327

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  • DOI: https://doi.org/10.2116/analsci.31.327

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