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
  • 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.
Products