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

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 cell...

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Autores principales:	Kenshi Yaginuma, Wataru Aoki, Natsuko Miura, Yuta Ohtani, Shunsuke Aburaya, Masato Kogawa, Yohei Nishikawa, Masahito Hosokawa, Haruko Takeyama, Mitsuyoshi Ueda
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2019
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/e4aeabe5ce914f16bea38ae6cd9902d6
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spelling	oai:doaj.org-article:e4aeabe5ce914f16bea38ae6cd9902d62021-12-02T15:09:33ZHigh-throughput identification of peptide agonists against GPCRs by co-culture of mammalian reporter cells and peptide-secreting yeast cells using droplet microfluidics10.1038/s41598-019-47388-x2045-2322https://doaj.org/article/e4aeabe5ce914f16bea38ae6cd9902d62019-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-019-47388-xhttps://doaj.org/toc/2045-2322Abstract 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.Kenshi YaginumaWataru AokiNatsuko MiuraYuta OhtaniShunsuke AburayaMasato KogawaYohei NishikawaMasahito HosokawaHaruko TakeyamaMitsuyoshi UedaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 9, Iss 1, Pp 1-11 (2019)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Kenshi Yaginuma Wataru Aoki Natsuko Miura Yuta Ohtani Shunsuke Aburaya Masato Kogawa Yohei Nishikawa Masahito Hosokawa Haruko Takeyama Mitsuyoshi Ueda High-throughput identification of peptide agonists against GPCRs by co-culture of mammalian reporter cells and peptide-secreting yeast cells using droplet microfluidics
description	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.
format	article
author	Kenshi Yaginuma Wataru Aoki Natsuko Miura Yuta Ohtani Shunsuke Aburaya Masato Kogawa Yohei Nishikawa Masahito Hosokawa Haruko Takeyama Mitsuyoshi Ueda
author_facet	Kenshi Yaginuma Wataru Aoki Natsuko Miura Yuta Ohtani Shunsuke Aburaya Masato Kogawa Yohei Nishikawa Masahito Hosokawa Haruko Takeyama Mitsuyoshi Ueda
author_sort	Kenshi Yaginuma
title	High-throughput identification of peptide agonists against GPCRs by co-culture of mammalian reporter cells and peptide-secreting yeast cells using droplet microfluidics
title_short	High-throughput identification of peptide agonists against GPCRs by co-culture of mammalian reporter cells and peptide-secreting yeast cells using droplet microfluidics
title_full	High-throughput identification of peptide agonists against GPCRs by co-culture of mammalian reporter cells and peptide-secreting yeast cells using droplet microfluidics
title_fullStr	High-throughput identification of peptide agonists against GPCRs by co-culture of mammalian reporter cells and peptide-secreting yeast cells using droplet microfluidics
title_full_unstemmed	High-throughput identification of peptide agonists against GPCRs by co-culture of mammalian reporter cells and peptide-secreting yeast cells using droplet microfluidics
title_sort	high-throughput identification of peptide agonists against gpcrs by co-culture of mammalian reporter cells and peptide-secreting yeast cells using droplet microfluidics
publisher	Nature Portfolio
publishDate	2019
url	https://doaj.org/article/e4aeabe5ce914f16bea38ae6cd9902d6
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High-throughput identification of peptide agonists against GPCRs by co-culture of mammalian reporter cells and peptide-secreting yeast cells using droplet microfluidics

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