5D3E

Crystal structure of human 14-3-3 gamma in complex with CFTR R-domain peptide pS768-pS795

Summary for 5D3E

• Entry DOI: 10.2210/pdb5d3e/pdb
• Descriptor: 14-3-3 protein gamma, Cystic fibrosis transmembrane conductance regulator (3 entities in total)
• Functional Keywords: protein-peptide complex, phosphorylation, tandem binding, signaling protein
• Biological source: Homo sapiens (Human); More
• Cellular location: Cytoplasm : P61981; Early endosome membrane ; Multi-pass membrane protein : P13569
• Total number of polymer chains: 9
• Total formula weight: 180433.27

Authors

Stevers, L.M.,Leysen, S.F.R.,Ottmann, C. (deposition date: 2015-08-06, release date: 2016-03-16, Last modification date: 2024-10-16)

Primary citation

Stevers, L.M.,Lam, C.V.,Leysen, S.F.,Meijer, F.A.,van Scheppingen, D.S.,de Vries, R.M.,Carlile, G.W.,Milroy, L.G.,Thomas, D.Y.,Brunsveld, L.,Ottmann, C.
Characterization and small-molecule stabilization of the multisite tandem binding between 14-3-3 and the R domain of CFTR.
Proc.Natl.Acad.Sci.USA, 113:E1152-E1161, 2016

PubMed Abstract: Cystic fibrosis is a fatal genetic disease, most frequently caused by the retention of the CFTR (cystic fibrosis transmembrane conductance regulator) mutant protein in the endoplasmic reticulum (ER). The binding of the 14-3-3 protein to the CFTR regulatory (R) domain has been found to enhance CFTR trafficking to the plasma membrane. To define the mechanism of action of this protein-protein interaction, we have examined the interaction in vitro. The disordered multiphosphorylated R domain contains nine different 14-3-3 binding motifs. Furthermore, the 14-3-3 protein forms a dimer containing two amphipathic grooves that can potentially bind these phosphorylated motifs. This results in a number of possible binding mechanisms between these two proteins. Using multiple biochemical assays and crystal structures, we show that the interaction between them is governed by two binding sites: The key binding site of CFTR (pS768) occupies one groove of the 14-3-3 dimer, and a weaker, secondary binding site occupies the other binding groove. We show that fusicoccin-A, a natural-product tool compound used in studies of 14-3-3 biology, can stabilize the interaction between 14-3-3 and CFTR by selectively interacting with a secondary binding motif of CFTR (pS753). The stabilization of this interaction stimulates the trafficking of mutant CFTR to the plasma membrane. This definition of the druggability of the 14-3-3-CFTR interface might offer an approach for cystic fibrosis therapeutics.

PubMed: 26888287
DOI: 10.1073/pnas.1516631113

Experimental method

X-RAY DIFFRACTION (2.75 Å)