Transcriptomics

Dataset Information

4

Characterization of RA839, a non-covalent small-molecule binder to Keap1 and selective activator of Nrf2 signalling


ABSTRACT: The activation of the transcription factor NF-E2-related factor 2 (Nrf2) maintains cellular homeostasis in response to oxidative stress by the regulation of multiple cytoprotective genes. Without stressors the activity of Nrf2 is inhibited by its interaction with the kelch-like ECH-associated protein 1 (Keap1). Here, we describe RA839, a small molecule that binds non-covalently to the Nrf2-interacting kelch domain of Keap1 with a Kd of approximately 6 µM, as demonstrated by X-ray co-crystallization and isothermal titration calorimetry. Whole-genome DNA arrays showed that at 10 µM RA839 significantly regulated 105 genes in bone marrow-derived macrophages. Canonical pathway mapping of these genes revealed an activation of pathways linked with Nrf2 signalling. These pathways were also activated after the activation of Nrf2 by the silencing of Keap1 expression. RA839 regulated only two genes in Nrf2 knockout macrophages. Similar to the activation of Nrf2 by either silencing of Keap1 expression or by the reactive compound CDDO-Me, RA839 prevented the induction of both inducible nitric oxide synthase expression and nitric oxide release in response to lipopolysaccharides in macrophages. In mice RA839 acutely induced Nrf2-target gene expression in liver. RA839 is a selective inhibitor of the Keap1/Nrf2 interaction and a useful tool compound to study the biology of Nrf2. Gene expression profile of bone marrow derived murine macrophages (BMDM) from Nrf2+/+ or Nrf2-/- mice treated with RA838, siKeap1-1 or siKeap1-2 were compared to untreated DMSO control or siControl. Four biological replicates were used for each sample group.

REANALYSED by: E-GEOD-71695

ORGANISM(S): Mus musculus  

SUBMITTER: Dieter Schmoll   Angelika Winkel  Daniel Margerie 

PROVIDER: E-GEOD-71695 | ArrayExpress | 2015-10-31

SECONDARY ACCESSION(S): GSE71695PRJNA291845

REPOSITORIES: GEO, ArrayExpress

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Publications

Characterization of RA839, a Noncovalent Small Molecule Binder to Keap1 and Selective Activator of Nrf2 Signaling.

Winkel Angelika F AF   Engel Christian K CK   Margerie Daniel D   Kannt Aimo A   Szillat Hauke H   Glombik Heiner H   Kallus Christopher C   Ruf Sven S   Güssregen Stefan S   Riedel Jens J   Herling Andreas W AW   von Knethen Andreas A   Weigert Andreas A   Brüne Bernhard B   Schmoll Dieter D  

The Journal of biological chemistry 20151012 47


The activation of the transcription factor NF-E2-related factor 2 (Nrf2) maintains cellular homeostasis in response to oxidative stress by the regulation of multiple cytoprotective genes. Without stressors, the activity of Nrf2 is inhibited by its interaction with the Keap1 (kelch-like ECH-associated protein 1). Here, we describe (3S)-1-[4-[(2,3,5,6-tetramethylphenyl) sulfonylamino]-1-naphthyl]pyrrolidine-3-carboxylic acid (RA839), a small molecule that binds noncovalently to the Nrf2-interactin  ...[more]

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