Discovery of Dimer-Dependent Aminoacrylamide Molecular Glues for 14–3–3 Protein–Protein Interactions

TL;DR

Researchers discovered a new type of molecular glue that stabilizes interactions involving 14–3–3 proteins, which are linked to various diseases.

Contribution

The study introduces a novel aminoacrylamide molecular glue that acts at the 14–3–3 dimer interface without targeting cysteine.

Findings

Aminoacrylamide 7 requires both a basic amine and acrylamide group for activity.

Compound 7 has a cysteine-independent mechanism of action.

Activity of 7 depends on 14–3–3 dimerization and affects interactions with ERα, LRRK2, and AHA2.

Abstract

Molecular glues (MGs) offer a promising strategy for
stabilizing
protein–protein interactions (PPIs), particularly within the
14–3–3 protein family, which regulates diverse cellular
processes and is implicated in many disease pathways. This study reports
on the discovery of an aminoacrylamide MG (7) for 14–3–3
PPIs. Structure–activity relationship analysis using a fluorescence
polarization (FP) assay revealed that both a basic amine and acrylamide
moiety are essential for activity. However, further investigation
using FP, mass spectrometry, and a thermal shift assay revealed that 7 has a cysteine-independent mode of action, distinguishing
it from other covalent 14–3–3 MGs. Furthermore, its
activity is reliant on 14–3–3 dimerization suggesting
that it targets the 14–3–3 dimer interface. Aminoacrylamide 7 differentially affected interactions with ERα, LRRK2,
and AHA2, suggesting that 14–3–3 dimerization plays
an important role in 14–3–3 client recognition. These
findings further validate the 14–3–3 dimer interface
as a novel MG target and underscore the complexities of 14–3–3
molecular recognition and small-molecule modulation.

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