Direct visualization of glucagon-like peptide-1 secretion by fluorescent fusion proteins

Tsuzuki, Atsushi; Fujioka, Yoichiro; Yoshida, Aiko; Kashiwagi, Sayaka; Amano, Maho; Hira, Tohru; Nakamura, Akinobu; Miyoshi, Hideaki; Atsumi, Tatsuya; Ohba, Yusuke

Direct visualization of glucagon-like peptide-1 secretion by fluorescent fusion proteins

Tsuzuki, Atsushi; Fujioka, Yoichiro; Yoshida, Aiko et al.

Journal of diabetes investigation, 2022, 13(7), 1134-1139

Permalink : https://hdl.handle.net/2115/86630

このアイテムのアクセス数:477件（2026-08-19 00:28 集計）

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https://doi.org/10.1111/jdi.13800			37

論文情報

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アクセス権	open access
URI	https://hdl.handle.net/2115/86630
タイトル	Direct visualization of glucagon-like peptide-1 secretion by fluorescent fusion proteins
著者	Tsuzuki, Atsushi ; Fujioka, Yoichiro ; Yoshida, Aiko ; Kashiwagi, Sayaka ; Amano, Maho ; Hira, Tohru ; Nakamura, Akinobu ; Miyoshi, Hideaki ; Atsumi, Tatsuya ; Ohba, Yusuke
著者名 Tsuzuki, Atsushi
e-Rad_Researcher 70597492 著者名 Fujioka, Yoichiro
e-Rad_Researcher 70831288 著者名 Yoshida, Aiko
e-Rad_Researcher 60880002 著者名 Kashiwagi, Sayaka
e-Rad_Researcher 80365808 著者名 Amano, Maho
e-Rad_Researcher 10396301 著者名 Hira, Tohru
e-Rad_Researcher 70552420 著者名 Nakamura, Akinobu
e-Rad_Researcher 30360902 著者名 Miyoshi, Hideaki
e-Rad_Researcher 20301905 著者名 Atsumi, Tatsuya
e-Rad_Researcher 30333503 著者名 Ohba, Yusuke
言語	英語
キーワード	Exocytosis
	Glucagon-like peptide-1
	Total internal reflection fluorescence microscope
発行日	2022-07
出版者	John Wiley & Sons
収録物名	Journal of diabetes investigation
巻(号)	13(7)
ページ	1134-1139
抄録	Live-cell imaging with fluorescent proteins (FPs) is a powerful tool for investigating the exocytosis processes of hormones. However, the secretion process of glucagon-like peptide-1 (GLP-1) has not been visualized by FPs, which might be because tagging FPs inhibits GLP-1 synthesis through the post-translational processing from proglucagon. Here, we have developed FP-tagged GLP-1 by inserting FPs into the middle of GLP-1 and adding the proglucagon signal peptide. Confocal imaging confirmed that GLP-1 fused to FPs with high folding efficiency showed granular structure, in which secretory vesicle markers colocalized. The fluorescence intensity of FP in the culture supernatant from cells treated with KCl or forskolin was significantly increased compared with those from untreated cells. Furthermore, FP-tagged GLP-1 enables direct visualization of stimulation-dependent exocytosis of GLP-1 at a single granule resolution with total internal reflection fluorescence microscopy. FP-tagged GLP-1 might facilitate the screening of GLP-1 secretagogues and the discovery of new antidiabetic drugs.
資源タイプ	学術雑誌論文
出版タイプ	Version of Record
PISSN	2040-1116
関連情報 (isIdenticalTo)	出版社版DOI https://doi.org/10.1111/jdi.13800