3ZGD

crystal structure of a KEAP1 mutant

Summary for 3ZGD

• Entry DOI: 10.2210/pdb3zgd/pdb
• Related: 3ZGC
• Descriptor: KELCH-LIKE ECH-ASSOCIATED PROTEIN 1, ACETATE ION, SODIUM ION, ... (4 entities in total)
• Functional Keywords: transcription, crystal contact engineering
• Biological source: HOMO SAPIENS (HUMAN)
• Cellular location: Cytoplasm: Q14145
• Total number of polymer chains: 2
• Total formula weight: 68295.13

Authors

Hoerer, S.,Reinert, D.,Ostmann, K.,Hoevels, Y.,Nar, H. (deposition date: 2012-12-17, release date: 2013-06-12, Last modification date: 2024-11-20)

Primary citation

Horer, S.,Reinert, D.,Ostmann, K.,Hoevels, Y.,Nar, H.
Crystal-Contact Engineering to Obtain a Crystal Form of the Kelch Domain of Human Keap1 Suitable for Ligand-Soaking Experiments.
Acta Crystallogr.,Sect.F, 69:592-, 2013

PubMed Abstract: Keap1 is a substrate adaptor protein for a Cul3-dependent ubiquitin ligase complex and plays an important role in the cellular response to oxidative stress. It binds Nrf2 with its Kelch domain and thus triggers the ubiquitinylation and degradation of Nrf2. Oxidative stress prevents the degradation of Nrf2 and leads to the activation of cytoprotective genes. Therefore, Keap1 is an attractive drug target in inflammatory diseases. The support of a medicinal chemistry effort by structural research requires a robust crystallization system in which the crystals are preferably suited for performing soaking experiments. This facilitates the generation of protein-ligand complexes in a routine and high-throughput manner. The structure of human Keap1 has been described previously. In this crystal form, however, the binding site for Nrf2 was blocked by a crystal contact. This interaction was analysed and mutations were introduced to disrupt this crystal contact. One double mutation (E540A/E542A) crystallized in a new crystal form in which the binding site for Nrf2 was not blocked and was accessible to small-molecule ligands. The crystal structures of the apo form of the mutated Keap1 Kelch domain (1.98 Å resolution) and of the complex with an Nrf2-derived peptide obtained by soaking (2.20 Å resolution) are reported.

PubMed: 23722832
DOI: 10.1107/S174430911301124X

Experimental method

X-RAY DIFFRACTION (1.98 Å)