Drug-Repositioning Screening for Keap1-Nrf2 Binding Inhibitors using Fluorescence Correlation Spectroscopy

Abstract The Kelch-like ECH-associating protein 1 (Keap1)-nuclear factor erythroid 2-related factor 2 (Nrf2)-antioxidant response element (ARE) signaling pathway is the major regulator of cytoprotective responses to oxidative and electrophilic stress. The Cul3/Keap1 E3 ubiquitin ligase complex inter...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Yuki Yoshizaki, Takayasu Mori, Mari Ishigami-Yuasa, Eriko Kikuchi, Daiei Takahashi, Moko Zeniya, Naohiro Nomura, Yutaro Mori, Yuya Araki, Fumiaki Ando, Shintaro Mandai, Yuri Kasagi, Yohei Arai, Emi Sasaki, Sayaka Yoshida, Hiroyuki Kagechika, Tatemitsu Rai, Shinichi Uchida, Eisei Sohara
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2017
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/9033bda222a94a88a5557b6bef9e8da0
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:9033bda222a94a88a5557b6bef9e8da0
record_format	dspace
spelling	oai:doaj.org-article:9033bda222a94a88a5557b6bef9e8da02021-12-02T16:07:46ZDrug-Repositioning Screening for Keap1-Nrf2 Binding Inhibitors using Fluorescence Correlation Spectroscopy10.1038/s41598-017-04233-32045-2322https://doaj.org/article/9033bda222a94a88a5557b6bef9e8da02017-06-01T00:00:00Zhttps://doi.org/10.1038/s41598-017-04233-3https://doaj.org/toc/2045-2322Abstract The Kelch-like ECH-associating protein 1 (Keap1)-nuclear factor erythroid 2-related factor 2 (Nrf2)-antioxidant response element (ARE) signaling pathway is the major regulator of cytoprotective responses to oxidative and electrophilic stress. The Cul3/Keap1 E3 ubiquitin ligase complex interacts with Nrf2, leading to Nrf2 ubiquitination and degradation. In this study, we focused on the disruption of the Keap1-Nrf2 interaction to upregulate Nrf2 expression and the transcription of ARE-controlled cytoprotective oxidative stress response enzymes, such as HO-1. We completed a drug-repositioning screening for inhibitors of Keap1-Nrf2 protein-protein interactions using a newly established fluorescence correlation spectroscopy (FCS) screening system. The binding reaction between Nrf2 and Keap1 was successfully detected with a KD of 2.6 μM using our FCS system. The initial screening of 1,633 drugs resulted in 12 candidate drugs. Among them, 2 drugs significantly increased Nrf2 protein levels in HepG2 cells. These two promising drugs also upregulated ARE gene promoter activity and increased HO-1 mRNA expression, which confirms their ability to dissociate Nrf2 and Keap1. Thus, drug-repositioning screening for Keap1-Nrf2 binding inhibitors using FCS enabled us to find two promising known drugs that can induce the activation of the Nrf2-ARE pathway.Yuki YoshizakiTakayasu MoriMari Ishigami-YuasaEriko KikuchiDaiei TakahashiMoko ZeniyaNaohiro NomuraYutaro MoriYuya ArakiFumiaki AndoShintaro MandaiYuri KasagiYohei AraiEmi SasakiSayaka YoshidaHiroyuki KagechikaTatemitsu RaiShinichi UchidaEisei SoharaNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 7, Iss 1, Pp 1-10 (2017)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Yuki Yoshizaki Takayasu Mori Mari Ishigami-Yuasa Eriko Kikuchi Daiei Takahashi Moko Zeniya Naohiro Nomura Yutaro Mori Yuya Araki Fumiaki Ando Shintaro Mandai Yuri Kasagi Yohei Arai Emi Sasaki Sayaka Yoshida Hiroyuki Kagechika Tatemitsu Rai Shinichi Uchida Eisei Sohara Drug-Repositioning Screening for Keap1-Nrf2 Binding Inhibitors using Fluorescence Correlation Spectroscopy
description	Abstract The Kelch-like ECH-associating protein 1 (Keap1)-nuclear factor erythroid 2-related factor 2 (Nrf2)-antioxidant response element (ARE) signaling pathway is the major regulator of cytoprotective responses to oxidative and electrophilic stress. The Cul3/Keap1 E3 ubiquitin ligase complex interacts with Nrf2, leading to Nrf2 ubiquitination and degradation. In this study, we focused on the disruption of the Keap1-Nrf2 interaction to upregulate Nrf2 expression and the transcription of ARE-controlled cytoprotective oxidative stress response enzymes, such as HO-1. We completed a drug-repositioning screening for inhibitors of Keap1-Nrf2 protein-protein interactions using a newly established fluorescence correlation spectroscopy (FCS) screening system. The binding reaction between Nrf2 and Keap1 was successfully detected with a KD of 2.6 μM using our FCS system. The initial screening of 1,633 drugs resulted in 12 candidate drugs. Among them, 2 drugs significantly increased Nrf2 protein levels in HepG2 cells. These two promising drugs also upregulated ARE gene promoter activity and increased HO-1 mRNA expression, which confirms their ability to dissociate Nrf2 and Keap1. Thus, drug-repositioning screening for Keap1-Nrf2 binding inhibitors using FCS enabled us to find two promising known drugs that can induce the activation of the Nrf2-ARE pathway.
format	article
author	Yuki Yoshizaki Takayasu Mori Mari Ishigami-Yuasa Eriko Kikuchi Daiei Takahashi Moko Zeniya Naohiro Nomura Yutaro Mori Yuya Araki Fumiaki Ando Shintaro Mandai Yuri Kasagi Yohei Arai Emi Sasaki Sayaka Yoshida Hiroyuki Kagechika Tatemitsu Rai Shinichi Uchida Eisei Sohara
author_facet	Yuki Yoshizaki Takayasu Mori Mari Ishigami-Yuasa Eriko Kikuchi Daiei Takahashi Moko Zeniya Naohiro Nomura Yutaro Mori Yuya Araki Fumiaki Ando Shintaro Mandai Yuri Kasagi Yohei Arai Emi Sasaki Sayaka Yoshida Hiroyuki Kagechika Tatemitsu Rai Shinichi Uchida Eisei Sohara
author_sort	Yuki Yoshizaki
title	Drug-Repositioning Screening for Keap1-Nrf2 Binding Inhibitors using Fluorescence Correlation Spectroscopy
title_short	Drug-Repositioning Screening for Keap1-Nrf2 Binding Inhibitors using Fluorescence Correlation Spectroscopy
title_full	Drug-Repositioning Screening for Keap1-Nrf2 Binding Inhibitors using Fluorescence Correlation Spectroscopy
title_fullStr	Drug-Repositioning Screening for Keap1-Nrf2 Binding Inhibitors using Fluorescence Correlation Spectroscopy
title_full_unstemmed	Drug-Repositioning Screening for Keap1-Nrf2 Binding Inhibitors using Fluorescence Correlation Spectroscopy
title_sort	drug-repositioning screening for keap1-nrf2 binding inhibitors using fluorescence correlation spectroscopy
publisher	Nature Portfolio
publishDate	2017
url	https://doaj.org/article/9033bda222a94a88a5557b6bef9e8da0
work_keys_str_mv	AT yukiyoshizaki drugrepositioningscreeningforkeap1nrf2bindinginhibitorsusingfluorescencecorrelationspectroscopy AT takayasumori drugrepositioningscreeningforkeap1nrf2bindinginhibitorsusingfluorescencecorrelationspectroscopy AT mariishigamiyuasa drugrepositioningscreeningforkeap1nrf2bindinginhibitorsusingfluorescencecorrelationspectroscopy AT erikokikuchi drugrepositioningscreeningforkeap1nrf2bindinginhibitorsusingfluorescencecorrelationspectroscopy AT daieitakahashi drugrepositioningscreeningforkeap1nrf2bindinginhibitorsusingfluorescencecorrelationspectroscopy AT mokozeniya drugrepositioningscreeningforkeap1nrf2bindinginhibitorsusingfluorescencecorrelationspectroscopy AT naohironomura drugrepositioningscreeningforkeap1nrf2bindinginhibitorsusingfluorescencecorrelationspectroscopy AT yutaromori drugrepositioningscreeningforkeap1nrf2bindinginhibitorsusingfluorescencecorrelationspectroscopy AT yuyaaraki drugrepositioningscreeningforkeap1nrf2bindinginhibitorsusingfluorescencecorrelationspectroscopy AT fumiakiando drugrepositioningscreeningforkeap1nrf2bindinginhibitorsusingfluorescencecorrelationspectroscopy AT shintaromandai drugrepositioningscreeningforkeap1nrf2bindinginhibitorsusingfluorescencecorrelationspectroscopy AT yurikasagi drugrepositioningscreeningforkeap1nrf2bindinginhibitorsusingfluorescencecorrelationspectroscopy AT yoheiarai drugrepositioningscreeningforkeap1nrf2bindinginhibitorsusingfluorescencecorrelationspectroscopy AT emisasaki drugrepositioningscreeningforkeap1nrf2bindinginhibitorsusingfluorescencecorrelationspectroscopy AT sayakayoshida drugrepositioningscreeningforkeap1nrf2bindinginhibitorsusingfluorescencecorrelationspectroscopy AT hiroyukikagechika drugrepositioningscreeningforkeap1nrf2bindinginhibitorsusingfluorescencecorrelationspectroscopy AT tatemitsurai drugrepositioningscreeningforkeap1nrf2bindinginhibitorsusingfluorescencecorrelationspectroscopy AT shinichiuchida drugrepositioningscreeningforkeap1nrf2bindinginhibitorsusingfluorescencecorrelationspectroscopy AT eiseisohara drugrepositioningscreeningforkeap1nrf2bindinginhibitorsusingfluorescencecorrelationspectroscopy
_version_	1718384730283442176

Drug-Repositioning Screening for Keap1-Nrf2 Binding Inhibitors using Fluorescence Correlation Spectroscopy

Ejemplares similares