Fragment-Based Interrogation of the 14-3-3/TAZ Protein-Protein Interaction.

Area of Science:

• Biochemistry
• Structural Biology
• Drug Discovery

Background:

• Developing molecular glues to modulate protein-protein interactions (PPIs) is challenging.
• The 14-3-3 protein family plays crucial roles in various cellular signaling pathways.
• Targeting 14-3-3 mediated PPIs presents therapeutic opportunities.

Purpose of the Study:

• To identify chemical starting points for molecular glue development targeting 14-3-3 protein interactions.
• To characterize a novel allosteric stabilizer of the 14-3-3/TAZ peptide complex.

Main Methods:

• Fragment screening was employed to identify potential binders.
• Differential scanning fluorimetry and microscale thermophoresis were used to assess stabilization.
• Molecular dynamics, nuclear magnetic resonance, and X-ray crystallography were utilized to determine the binding site.

Main Results:

• A fragment was identified that preferentially binds to the 14-3-3/TAZ peptide complex.
• The fragment demonstrated moderate stabilization of the complex.
• Structural studies revealed an allosteric binding site between helices 8 and 9 of 14-3-3, distinct from the peptide interface.

Conclusions:

• This study presents the first example of an allosteric stabilizer for 14-3-3 protein-protein interactions.
• The identified fragment serves as a valuable chemical starting point for developing novel molecular glues.
• This work opens new avenues for modulating 14-3-3 mediated signaling pathways through allosteric stabilization.