2. Academic Validation
  3. The isoquinoline PRL-295 increases the thermostability of Keap1 and disrupts its interaction with Nrf2

The isoquinoline PRL-295 increases the thermostability of Keap1 and disrupts its interaction with Nrf2

  • iScience. 2021 Dec 27;25(1):103703. doi: 10.1016/j.isci.2021.103703.
Sharadha Dayalan Naidu 1,  Takafumi Suzuki 2,  Dina Dikovskaya 1,  Elena V Knatko 1,  Maureen Higgins 1,  Miu Sato 2,  Miroslav Novak 1,  José A Villegas 3,  Terry W Moore 3,  Masayuki Yamamoto 2,  Albena T Dinkova-Kostova 1 4
Affiliations

Affiliations

  • 1 Division of Cellular Medicine, School of Medicine, Jacqui Wood Cancer Centre, Ninewells Hospital and Medical School, University of Dundee, James Arnott Drive, Dundee, Scotland DD1 9SY, UK.
  • 2 Department of Medical Biochemistry, Tohoku University Graduate School of Medicine, Sendai, Japan.
  • 3 Department of Pharmaceutical Sciences, College of Pharmacy, University of Illinois Cancer Center, University of Illinois at Chicago, Chicago, IL 60607, USA.
  • 4 Department of Pharmacology and Molecular Sciences and Department of Medicine, Johns Hopkins University School of Medicine, Baltimore, MD 21205, USA.
PMID: 35036882 DOI: 10.1016/j.isci.2021.103703
Abstract

Transcription factor Nrf2 and its negative regulator Keap1 orchestrate a cytoprotective response against oxidative, metabolic, and inflammatory stress. Keap1 is a drug target, with several small molecules in drug development. Here, we show that the isoquinoline PRL-295 increased Keap1 thermostability in lysates from cells expressing fluorescently tagged Keap1. The thermostability of endogenous Keap1 also increased in intact cells and murine liver following PRL-295 treatment. Fluorescence Lifetime Imaging-Förster Resonance Energy Transfer (FLIM-FRET) experiments in cells co-expressing sfGFP-Nrf2 and Keap1-mCherry further showed that PRL-295 prolonged the donor fluorescence lifetime, indicating disruption of the Keap1-Nrf2 protein complex. Orally administered PRL-295 to mice activated the Nrf2transcriptional target NAD(P)H:quinone oxidoreductase 1 (NQO1) in liver and decreased the levels of plasma alanine aminotransferase and aspartate aminotransferase upon acetaminophen-induced hepatic injury. Thus, PRL-295 engages the Keap1 protein target in cells and in vivo, disrupting its interaction with Nrf2, leading to activation of Nrf2-dependent transcription and hepatocellular protection.

Keywords

Biochemistry; Biological sciences; Molecular interaction.

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