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Published on: August 27, 2021
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Lysine-Targeted Reversible Covalent Ligand Discovery for Proteins via Phage Display
Mengmeng Zheng1, Fa-Jie Chen1, Kaicheng Li1
1Department of Chemistry, Merkert Chemistry Center, Boston College, 2609 Beacon Street, Chestnut Hill, Massachusetts 02467, United States.
Journal of the American Chemical Society
|August 17, 2022
Summary
Phage display was used to create novel reversible covalent inhibitors. This new method successfully identified potent peptide ligands for bacterial and viral proteins, offering a versatile platform for drug discovery.
Area of Science:
- Medicinal Chemistry
- Biotechnology
- Drug Discovery
Background:
- Reversible covalent bond formation is a powerful strategy for designing potent protein inhibitors, exemplified by drugs like bortezomib and voxelotor.
- Rational design of such inhibitors is challenging, even with existing noncovalent scaffolds.
Purpose of the Study:
- To develop a phage display platform for generating reversible covalent inhibitors.
- To incorporate 2-acetylphenylboronic acid (APBA) as a warhead for reversible covalent binding to lysine residues.
Main Methods:
- Construction of chemically modified phage libraries (linear and cyclic) presenting APBA.
- Screening of APBA-presenting phage libraries against Sortase A (Staphylococcus aureus) and SARS-CoV-2 spike protein.
- Structure-activity relationship studies to confirm the role of the APBA warhead.
Main Results:
- Identification of peptide ligands with single-digit micromolar potency and high specificity for both target proteins.
- Demonstration of live-cell sortase inhibition and sensitive spike protein detection using the identified ligands.
- Unambiguous evidence from structure-activity studies highlighting the benefit of the APBA warhead for protein binding.
Conclusions:
- This study establishes phage display as a viable method for developing reversible covalent inhibitors against therapeutic targets.
- The APBA-presenting phage display platform is broadly applicable for discovering inhibitors for various proteins, including those involved in protein-protein interactions.
