TY - JOUR
T1 - The isoquinoline PRL-295 increases the thermostability of Keap1 and disrupts its interaction with Nrf2
AU - Dayalan Naidu, Sharadha
AU - Suzuki, Takafumi
AU - Dikovskaya, Dina
AU - Knatko, Elena V.
AU - Higgins, Maureen
AU - Sato, Miu
AU - Novak, Miroslav
AU - Villegas, José A.
AU - Moore, Terry W.
AU - Yamamoto, Masayuki
AU - Dinkova-Kostova, Albena T.
N1 - Funding Information:
We thank Phillip Lazzara for synthesizing PRL-295, the Biotechnology and Biological Sciences Research Council ( BB/L01923X/1 and BB/T508111/1 ), Tenovus Scotland ( T17/14 ), MEXT / JSPS KAKENHI ( 19H05649 , 19K07340 and 17KK0183 ), Cancer Research UK ( C20953 / A18644 ), and Reata Pharmaceuticals for financial support.
Publisher Copyright:
© 2022 The Authors
PY - 2022/1/21
Y1 - 2022/1/21
N2 - 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.
AB - 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.
KW - Biochemistry
KW - Biological sciences
KW - Molecular interaction
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U2 - 10.1016/j.isci.2021.103703
DO - 10.1016/j.isci.2021.103703
M3 - Article
AN - SCOPUS:85122539558
SN - 2589-0042
VL - 25
JO - iScience
JF - iScience
IS - 1
M1 - 103703
ER -