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Title: High-throughput identification of peptide agonists against GPCRs by co-culture of mammalian reporter cells and peptide-secreting yeast cells using droplet microfluidics
Authors: Yaginuma, Kenshi
Aoki, Wataru
Miura, Natsuko
Ohtani, Yuta
Aburaya, Shunsuke
Kogawa, Masato
Nishikawa, Yohei
Hosokawa, Masahito
Takeyama, Haruko
Ueda, Mitsuyoshi
Author's alias: 柳沼, 謙志
青木, 航
三浦, 夏子
大谷, 優太
油屋, 駿介
小川, 雅人
西川, 洋平
細川, 正人
竹山, 春子
植田, 充美
Keywords: Assay systems
High-throughput screening
Issue Date: 29-Jul-2019
Publisher: Springer Nature
Journal title: Scientific Reports
Volume: 9
Thesis number: 10920
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.
Description: ナノバイオ手法による創薬ターゲットGPCRの機能性リガンド探索のハイスループット化に成功 --ドロップレットマイクロ流体デバイスの活用--. 京都大学プレスリリース. 2019-08-02.
Rights: © The Author(s) 2019. This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.
URI: http://hdl.handle.net/2433/243246
DOI(Published Version): 10.1038/s41598-019-47388-x
PubMed ID: 31358824
Related Link: http://www.kyoto-u.ac.jp/ja/research/research_results/2019/190729_1.html
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