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

Sharadha Dayalan Naidu, Takafumi Suzuki, Dina Dikovskaya, Elena V. Knatko, Maureen Higgins, Miu Sato, Miroslav Novak, José A. Villegas, Terry W. Moore, Masayuki Yamamoto, Albena Dinkova-Kostova (Lead / Corresponding author)

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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 Nrf2-transcriptional 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.
Original languageEnglish
Article number103703
Number of pages19
Issue number1
Early online date27 Dec 2021
Publication statusPublished - 21 Jan 2022


  • Biochemistry
  • Biological sciences
  • Molecular interaction

ASJC Scopus subject areas

  • General


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