Yeast-based ligand assay system for detecting G protein-coupled receptor activation in water-in-oil droplets

Takashi Sakurai, Ryo Iizuka, Yasuyuki Tanigaki, Rui Sekine, Dong Hyun Yoon, Tetsushi Sekiguchi, Jun Ishii, Akihiko Kondo, Naoto Nemoto, Shuichi Shoji, Takashi Funatsu

Research output: Chapter in Book/Report/Conference proceedingConference contribution

1 Citation (Scopus)

Abstract

We present a yeast-based ligand assay system to detect activation of G protein-coupled receptors (GPCRs) in water-in-oil (W/O) droplets. A microfluidic device is used to encapsulate template DNA encoding peptides, a cell-free coupled transcription-translation system, and budding yeast cells expressing GPCR into W/O droplets. The yeast cells are genetically engineered to express green fluorescent protein (GFP) via their intracellular signaling pathway in response to ligand stimulation of GPCR. We successfully demonstrated that the synthesized peptide ligand activated the cognate GPCR in W/O droplets. This will be a new platform for identifying novel peptide agonists for mammalian GPCRs.

Original languageEnglish
Title of host publicationProceedings of the 16th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2012
PublisherChemical and Biological Microsystems Society
Pages1537-1539
Number of pages3
ISBN (Print)9780979806452
Publication statusPublished - 2012 Jan 1
Event16th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2012 - Okinawa, Japan
Duration: 2012 Oct 282012 Nov 1

Publication series

NameProceedings of the 16th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2012

Conference

Conference16th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2012
CountryJapan
CityOkinawa
Period12/10/2812/11/1

Keywords

  • Cell-free protein synthesis
  • Droplet
  • GPCR
  • Ligand assay
  • Yeast

ASJC Scopus subject areas

  • Chemical Engineering (miscellaneous)
  • Bioengineering

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  • Cite this

    Sakurai, T., Iizuka, R., Tanigaki, Y., Sekine, R., Yoon, D. H., Sekiguchi, T., Ishii, J., Kondo, A., Nemoto, N., Shoji, S., & Funatsu, T. (2012). Yeast-based ligand assay system for detecting G protein-coupled receptor activation in water-in-oil droplets. In Proceedings of the 16th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2012 (pp. 1537-1539). (Proceedings of the 16th International Conference on Miniaturized Systems for Chemistry and Life Sciences, MicroTAS 2012). Chemical and Biological Microsystems Society.