30ZX

Binary structure of 14-3-3sigma and interaction cassette phosphopeptide 2

Summary for 30ZX

• Entry DOI: 10.2210/pdb30zx/pdb
• Descriptor: 14-3-3 protein sigma, interaction cassette peptide 2, MAGNESIUM ION, ... (5 entities in total)
• Functional Keywords: hub protein 14-3-3, protein-protein interaction, stabilizer, peptide binding protein
• Biological source: Homo sapiens (human); More
• Total number of polymer chains: 2
• Total formula weight: 28280.01

Authors

Ni, Y.,Brusveld, L.,Ottmann, C.,Munoz Lasso, D. (deposition date: 2026-05-19, release date: 2026-09-02)

Primary citation

Munoz-Lasso, D.C.,Ni, Y.,Weber, G.,Eduati, F.,Beijersbergen, R.L.,Ottmann, C.,Brunsveld, L.
Molecular Glue and Phosphorylation-Dependent 14-3-3 Recruitment to p53 with an Engineered Interaction Cassette.
Acs Chem.Biol., 2026

PubMed Abstract: The tumor suppressor p53 is regulated by phosphorylation-dependent protein-protein interactions, including via binding to 14-3-3 adaptor proteins, which can tune p53 activity. Molecular glue (MG)-induced stabilization of 14-3-3/client interactions offers an attractive strategy to probe such networks, but cellular engagement is often constrained by context-dependent phosphorylation and interaction occupancy. Here, we engineered phosphorylation- and MG-dependent 14-3-3 interaction cassettes (IC1 and IC2) and fused them to p53 to promote recruitment of endogenous 14-3-3 proteins in human cells. Biochemical characterization establishes high-affinity binding of the phosphorylated cassettes to 14-3-3 and enhanced in vitro stabilization by the 14-3-3 molecular glue 3'-deacetylated fusicoccin-A (FC-A). In HEK293T cells, Flag-p53-IC1 and Flag-p53-IC2 co-immunoprecipitated native 14-3-3 proteins. Mutation of the cassette's phospho-accepting serine to alanine abolished binding, confirming phosphorylation dependent recruitment. Transcriptomic profiling of transiently transfected cells reveals cassette-dependent remodeling of a p53-associated gene expression landscape. Together, these results establish a modular, MG- and phosphorylation-dependent platform for engaging 14-3-3 in a p53 context and for evaluating how chemical stabilization translates from biochemical interaction control to cellular pathway-level readouts.

PubMed: 42631642
DOI: 10.1021/acschembio.6c00482

Experimental method

X-RAY DIFFRACTION (1.5 Å)