title: Structure of the BTB domain of Keap1 and its interaction with the triterpenoid antagonist CDDO.
abstract: The protein Keap1 is central to the regulation of the Nrf2-mediated cytoprotective response, and is increasingly recognized as an important target for therapeutic intervention in a range of diseases involving excessive oxidative stress and inflammation. The BTB domain of Keap1 plays key roles in sensing environmental electrophiles and in mediating interactions with the Cul3/Rbx1 E3 ubiquitin ligase system, and is believed to be the target for several small molecule covalent activators of the Nrf2 pathway. However, despite structural information being available for several BTB domains from related proteins, there have been no reported crystal structures of Keap1 BTB, and this has precluded a detailed understanding of its mechanism of action and interaction with antagonists. We report here the first structure of the BTB domain of Keap1, which is thought to contain the key cysteine residue responsible for interaction with electrophiles, as well as structures of the covalent complex with the antagonist CDDO/bardoxolone, and of the constitutively inactive C151W BTB mutant. In addition to providing the first structural confirmation of antagonist binding to Keap1 BTB, we also present biochemical evidence that adduction of Cys 151 by CDDO is capable of inhibiting the binding of Cul3 to Keap1, and discuss how this class of compound might exert Nrf2 activation through disruption of the BTB-Cul3 interface.
authors.affiliation: Astex Pharmaceuticals, Cambridge, United Kingdom.
authors.forename: Anne
authors.initials: A
authors.lastname: Cleasby
authors.affiliation: Astex Pharmaceuticals, Cambridge, United Kingdom.
authors.forename: Jeff
authors.initials: J
authors.lastname: Yon
authors.affiliation: Astex Pharmaceuticals, Cambridge, United Kingdom.
authors.forename: Philip J
authors.initials: PJ
authors.lastname: Day
authors.affiliation: Astex Pharmaceuticals, Cambridge, United Kingdom.
authors.forename: Caroline
authors.initials: C
authors.lastname: Richardson
authors.affiliation: Astex Pharmaceuticals, Cambridge, United Kingdom.
authors.forename: Ian J
authors.initials: IJ
authors.lastname: Tickle
authors.affiliation: Astex Pharmaceuticals, Cambridge, United Kingdom.
authors.forename: Pamela A
authors.initials: PA
authors.lastname: Williams
authors.affiliation: GlaxoSmithKline, King of Prussia, Pennsylvania, United States of America.
authors.forename: James F
authors.initials: JF
authors.lastname: Callahan
authors.affiliation: GlaxoSmithKline, Stevenage, United Kingdom.
authors.forename: Robin
authors.initials: R
authors.lastname: Carr
authors.affiliation: GlaxoSmithKline, Collegeville, Pennsylvania, United States of America.
authors.forename: Nestor
authors.initials: N
authors.lastname: Concha
authors.affiliation: GlaxoSmithKline, King of Prussia, Pennsylvania, United States of America.
authors.forename: Jeffrey K
authors.initials: JK
authors.lastname: Kerns
authors.affiliation: GlaxoSmithKline, Collegeville, Pennsylvania, United States of America.
authors.forename: Hongwei
authors.initials: H
authors.lastname: Qi
authors.affiliation: GlaxoSmithKline, King of Prussia, Pennsylvania, United States of America.
authors.forename: Thomas
authors.initials: T
authors.lastname: Sweitzer
authors.affiliation: GlaxoSmithKline, Collegeville, Pennsylvania, United States of America.
authors.forename: Paris
authors.initials: P
authors.lastname: Ward
authors.affiliation: Astex Pharmaceuticals, Cambridge, United Kingdom.
authors.forename: Thomas G
authors.initials: TG
authors.lastname: Davies
mesh_terms: D001665:Binding Sites; D006801:Humans; D007093:Imidazoles / Q000737:chemistry* / Q000494:pharmacology; D047908:Intracellular Signaling Peptides and Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000235:genetics; D000072019:Kelch-Like ECH-Associated Protein 1; D008958:Models, Molecular; D008968:Molecular Conformation; D009154:Mutation; D009828:Oleanolic Acid / Q000031:analogs & derivatives* / Q000737:chemistry / Q000494:pharmacology; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D013329:Structure-Activity Relationship
chemical_list: C472829:1-(2-cyano-3,12-dioxooleana-1,9-dien-28-oyl) imidazole; D007093:Imidazoles; D047908:Intracellular Signaling Peptides and Proteins; C431364:KEAP1 protein, human; D000072019:Kelch-Like ECH-Associated Protein 1; D009828:Oleanolic Acid