Design of Ligand-Operable Protein-Cages That Open Upon Specific Protein Binding
Eric J Lee1, Nika Gladkov1, Justin E Miller2
1Department of Chemistry and Biochemistry, UCLA, Los Angeles, California 90095, United States.
ACS Synthetic Biology
|December 22, 2023
Summary
We developed Ligand-Operable Cages (LOCs), designed protein cages that disassemble upon target protein binding. This enables controlled release of encapsulated molecules for advanced molecular delivery and detection applications.
Area of Science:
- Biotechnology
- Materials Science
- Molecular Biology
Background:
- Protein nanocages are useful for molecular delivery but lack controlled release mechanisms.
- Existing methods for nanocage opening are limited, hindering targeted applications.
- A modular system for protein-target-based nanocage disassembly is needed.
Purpose of the Study:
- To engineer a novel class of protein cages, Ligand-Operable Cages (LOCs), for controlled cargo release.
- To create a modular platform that responds to specific protein ligands for targeted delivery.
- To demonstrate a mechanism for triggered nanocage disassembly via protein-ligand interactions.
Main Methods:
- Designed protein cages (LOCs) with a core nanocage and a fused surface binding adaptor.
- Engineered steric hindrance between ligand binding and cage assembly for disassembly.
- Utilized natural and designed protein cages with various protein ligands and reporter readouts (fluorescence, luminescence).
Main Results:
- Demonstrated that target ligand binding induces LOC disassembly through mass action.
- Showcased the modularity of LOCs by reprogramming them to respond to different protein ligands.
- Successfully employed fluorescence unquenching and luminescence assays to report on cage disassembly.
Conclusions:
- LOCs provide a new platform for controlled molecular release triggered by specific protein targets.
- This technology advances targeted molecular delivery and detection capabilities.
- The modular design allows for versatile applications by altering the surface binder sequence.
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