Summary
This proposal focuses on the systematic discovery of selective small-molecule protein-protein interaction (PPI) stabilizers, using the hub protein 14-3-3σ as a model system to develop a toolkit for biological study and potential new drugs.
Systematic stabilization of specific protein-protein interactions
US · IL
NIH
grant
open
#nih-5R01GM147696-04
What they want
The project aims to develop a toolkit of selective, cell-active stabilizers for native 14-3-3/client PPIs. This will involve three main aims: 1) Screening for selective stabilizers of 14-3-3/phosphopeptide clients, using a native C38 residue on the 14-3-3σ isoform. 2) Optimizing 14-3-3/client stabilizers for cell-based activity, converting disulfides to cell-active electrophilic warheads and tuning selectivity for clients like CRAF kinase, estrogen receptor α (ERα), and FOXO1. 3) Designing PROTAC-based degraders of 14-3-3 clients, expanding PROTAC technology by using 14-3-3 as a scaffolding protein to link intrinsically disordered proteins (IDPs) to ubiquitin ligase, starting with ERα and then translating to IDPs like FOXO1.
Deliverables
- Tool kit of selective, cell-active stabilizers of native 14-3-3/client PPIs
- Proof-of-concept for the systematic discovery approach
- Optimized C38-bound fragments (with or without an electrophile) for target-selective PPI stabilization in cells
- PROTAC-based degraders of 14-3-3 clients
- Approaches broadly applicable to the 14-3-3 network and extendable to other native and nonnative PPIs
Technical requirements
- Screening for disulfide-bound fragments that stabilize 14-3-3/phosphopeptide complexes
- Utilization of native C38 residue on the 14-3-3σ isoform for screening
- Conversion of disulfides to cell-active electrophilic warheads
- Tuning the selectivity of 14-3-3σ/client stabilizers
- Design of PROTACs (bifunctional molecules that induce proximity between a ubiquitin ligase and a target)
- Use of 14-3-3 as a scaffolding protein to link intrinsically disordered proteins (IDPs) to ubiquitin ligase
