8RD1
HUWE1 WWE domain in complex with compound 4
Summary for 8RD1
Entry DOI | 10.2210/pdb8rd1/pdb |
Descriptor | E3 ubiquitin-protein ligase HUWE1, CHLORIDE ION, 2-(2-hydroxy-2-oxoethyl)-1,3-bis(oxidanylidene)isoindole-5-carboxylic acid, ... (4 entities in total) |
Functional Keywords | wwe domain, ligase |
Biological source | Homo sapiens (human) |
Total number of polymer chains | 4 |
Total formula weight | 43003.04 |
Authors | Muenzker, L.,Zak, K.M.,Boettcher, J. (deposition date: 2023-12-07, release date: 2024-07-31, Last modification date: 2024-08-07) |
Primary citation | Munzker, L.,Kimani, S.W.,Fowkes, M.M.,Dong, A.,Zheng, H.,Li, Y.,Dasovich, M.,Zak, K.M.,Leung, A.K.L.,Elkins, J.M.,Kessler, D.,Arrowsmith, C.H.,Halabelian, L.,Bottcher, J. A ligand discovery toolbox for the WWE domain family of human E3 ligases. Commun Biol, 7:901-901, 2024 Cited by PubMed Abstract: The WWE domain is a relatively under-researched domain found in twelve human proteins and characterized by a conserved tryptophan-tryptophan-glutamate (WWE) sequence motif. Six of these WWE domain-containing proteins also contain domains with E3 ubiquitin ligase activity. The general recognition of poly-ADP-ribosylated substrates by WWE domains suggests a potential avenue for development of Proteolysis-Targeting Chimeras (PROTACs). Here, we present novel crystal structures of the HUWE1, TRIP12, and DTX1 WWE domains in complex with PAR building blocks and their analogs, thus enabling a comprehensive analysis of the PAR binding site structural diversity. Furthermore, we introduce a versatile toolbox of biophysical and biochemical assays for the discovery and characterization of novel WWE domain binders, including fluorescence polarization-based PAR binding and displacement assays, N-NMR-based binding affinity assays and F-NMR-based competition assays. Through these assays, we have characterized the binding of monomeric iso-ADP-ribose (iso-ADPr) and its nucleotide analogs with the aforementioned WWE proteins. Finally, we have utilized the assay toolbox to screen a small molecule fragment library leading to the successful discovery of novel ligands targeting the HUWE1 WWE domain. PubMed: 39048679DOI: 10.1038/s42003-024-06584-w PDB entries with the same primary citation |
Experimental method | X-RAY DIFFRACTION (1.896 Å) |
Structure validation
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