9GLQ

Crystal structure of p73 tetramerisation domain in complex with darpins 1800

Summary for 9GLQ

• Entry DOI: 10.2210/pdb9glq/pdb
• Descriptor: Tumor protein p73, Darpins 1800, COBALT (II) ION, ... (5 entities in total)
• Functional Keywords: tp73, tetramerisation domain, darpins, structural genomics, structural genomics consortium, sgc, dna binding protein
• Biological source: Homo sapiens (human); More
• Total number of polymer chains: 3
• Total formula weight: 25494.63

Authors

Chaikuad, A.,Strubel, A.,Doetsch, V.,Knapp, S.,Structural Genomics Consortium (SGC) (deposition date: 2024-08-27, release date: 2025-01-22)

Primary citation

Munick, P.,Zielinski, J.,Strubel, A.,Gutfreund, N.,Dreier, B.,Schaefer, J.V.,Schafer, B.,Gebel, J.,Osterburg, C.,Chaikuad, A.,Knapp, S.,Pluckthun, A.,Dotsch, V.
DARPins as a novel tool to detect and degrade p73.
Cell Death Dis, 15:909-909, 2024

PubMed Abstract: The concept of Targeted Protein Degradation (TPD) has been introduced as an attractive alternative to the development of classical inhibitors. TPD can extend the range of proteins that can be pharmacologically targeted beyond the classical targets for small molecule inhibitors, as a binding pocket is required but its occupancy does not need to lead to inhibition. The method is based on either small molecules that simultaneously bind to a protein of interest and to a cellular E3 ligase and bring them in close proximity (molecular glue) or a bi-functional molecule synthesized from the chemical linkage of a target protein-specific small molecule and one that binds to an E3 ligase (Proteolysis Targeting Chimeras (PROTAC)). The further extension of this approach to bioPROTACs, in which a small protein-based binding module is fused directly to an E3 ligase or an E3 ligase adaptor protein, makes virtually all proteins amenable to targeted degradation, as this method eliminates the requirement for binding pockets for small molecules. Designed Ankyrin Repeat Proteins (DARPins) represent a very attractive class of small protein-based binding modules that can be used for the development of bioPTOTACS. Here we describe the characterization of two DARPins generated against the oligomerization domain and the SAM domain of the transcription factor p73, a member of the p53 protein family. The DARPins can be used for (isoform-)selective pulldown experiments both in cell culture as well as primary tissue lysates. We also demonstrate that they can be used for staining in cell culture experiments. Fusing them to the speckle type POZ protein (SPOP), an adaptor protein for cullin-3 E3 ligase complexes, yields highly selective and effective degraders. We demonstrate that selective degradation of the ΔNp73α isoform reactivates p53.

PubMed: 39695090
DOI: 10.1038/s41419-024-07304-2

Experimental method

X-RAY DIFFRACTION (2.1 Å)