High-throughput identification of peptide agonists against GPCRs by co-culture of mammalian reporter cells and peptide-secreting yeast cells using droplet microfluidics

Kenshi Yaginuma, Wataru Aoki, Natsuko Miura, Yuta Ohtani, Shunsuke Aburaya, Masato Kogawa, Yohei Nishikawa, Masahito Hosokawa, Haruko Takeyama, Mitsuyoshi Ueda

Research output: Contribution to journalArticle

Abstract

Since G-protein coupled receptors (GPCRs) are linked to various diseases, screening of functional ligands against GPCRs is vital for drug discovery. In the present study, we developed a high-throughput functional cell-based assay by combining human culture cells producing a GPCR, yeast cells secreting randomized peptide ligands, and a droplet microfluidic device. We constructed a reporter human cell line that emits fluorescence in response to the activation of human glucagon-like peptide-1 receptor (hGLP1R). We then constructed a yeast library secreting an agonist of hGLP1R or randomized peptide ligands. We demonstrated that high-throughput identification of functional ligands against hGLP1R could be performed by co-culturing the reporter cells and the yeast cells in droplets. We identified functional ligands, one of which had higher activity than that of an original sequence. The result suggests that our system could facilitate the discovery of functional peptide ligands of GPCRs.

Original languageEnglish
Article number10920
JournalScientific reports
Volume9
Issue number1
DOIs
Publication statusPublished - 2019 Dec 1

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Microfluidics
G-Protein-Coupled Receptors
Coculture Techniques
Yeasts
Ligands
Peptides
Lab-On-A-Chip Devices
Drug Discovery
Libraries
Cell Culture Techniques
Fluorescence
Cell Line
Glucagon-Like Peptide-1 Receptor

ASJC Scopus subject areas

  • General

Cite this

High-throughput identification of peptide agonists against GPCRs by co-culture of mammalian reporter cells and peptide-secreting yeast cells using droplet microfluidics. / Yaginuma, Kenshi; Aoki, Wataru; Miura, Natsuko; Ohtani, Yuta; Aburaya, Shunsuke; Kogawa, Masato; Nishikawa, Yohei; Hosokawa, Masahito; Takeyama, Haruko; Ueda, Mitsuyoshi.

In: Scientific reports, Vol. 9, No. 1, 10920, 01.12.2019.

Research output: Contribution to journalArticle

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AU - Ohtani, Yuta

AU - Aburaya, Shunsuke

AU - Kogawa, Masato

AU - Nishikawa, Yohei

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