¹H, ¹⁵N and ¹³C backbone resonance assignments of the Kelch domain of mouse Keap1.
Kelch-like ECH-associated Protein 1 (Keap1) is a multi-domain protein that functions as an inhibitor of the transcription factor nuclear factor E2-related factor 2 (Nrf2) in the cellular response to oxidative stress. Under normal conditions, Keap1 binds to Nrf2 via its C-terminal Kelch domain and the interaction ultimately leads to the ubiquitin-dependent degradation of Nrf2. It has been proposed that designing molecules to selectively disrupt the Keap1-Nrf2 interaction can be a potential therapeutic approach for enhancing the expression of cytoprotective genes. Here, we reported the (1)H, (13)C, and (15)N backbone chemical shift assignments of the Kelch domain of mouse Keap1. Further, unlabeled Nrf2 peptide containing the Kelch-binding motif was added to the (15)N-labeled Kelch sample. (1)H-(15)N HSQC spectra of the protein in the absence and presence of an equimolar concentration of the Nrf2 peptide were presented. A significant number of resonance signals were shifted upon addition of the peptide, confirming the protein-peptide interaction. The results here will not just facilitate the further studies of the binding between Keap1 and Nrf2, it will also be valuable for probing interactions between the Kelch domain and small molecules, as well as a growing list of protein targets that have been identified recently.
Department of Biochemistry, The University of Western Ontario, London, ON N6A 5C1, Canada.
Elio
E
Cino
Jingsong
J
Fan
Daiwen
D
Yang
Wing-Yiu
WY
Choy
D048868:Adaptor Proteins, Signal Transducing / Q000737:chemistry*; D000595:Amino Acid Sequence; D000818:Animals; D002247:Carbon Isotopes; D003598:Cytoskeletal Proteins / Q000737:chemistry*; D000072019:Kelch-Like ECH-Associated Protein 1; D051379:Mice; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009587:Nitrogen Isotopes; D019906:Nuclear Magnetic Resonance, Biomolecular; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011522:Protons
D048868:Adaptor Proteins, Signal Transducing; D002247:Carbon Isotopes; D003598:Cytoskeletal Proteins; C117301:Keap1 protein, mouse; D000072019:Kelch-Like ECH-Associated Protein 1; D009587:Nitrogen Isotopes; D011522:Protons