Asparaginyl Endopeptidase-Mediated Peptide Cyclization for Phage Display

Area of Science:

• Biochemistry
• Molecular Biology
• Drug Discovery

Background:

• Peptide cyclization is crucial for enhancing peptide stability and biological activity.
• Enzymatic methods offer precise and efficient routes for peptide modification.
• Targeting protein-protein interactions, like those involving TEAD4, is a key area in drug development.

Purpose of the Study:

• To develop a novel enzymatic strategy for asparaginyl endopeptidase-mediated peptide cyclization.
• To identify high-affinity macrocyclic peptide ligands for TEAD4 using the developed platform.
• To demonstrate the efficacy of the platform in discovering potent peptide therapeutics.

Main Methods:

• Enzymatic peptide cyclization using chloroacetylated recognition sequences and Cysteine residues with OaAEP1.
• Phage display technology for screening and identifying peptide ligands.
• Biacore analysis to determine binding affinity (KD) of identified peptides to TEAD4.

Main Results:

• Successful intramolecular cyclization of peptides mediated by OaAEP1.
• Identification of nanomolar macrocyclic peptide ligands targeting TEAD4.
• A bicyclic peptide ligand demonstrated a KD of 139 nM, showing 16-fold improvement over its linear analog.

Conclusions:

• The enzymatic strategy enables efficient generation of macrocyclic peptides.
• The platform is effective for discovering high-affinity peptide ligands against challenging targets like TEAD4.
• This approach holds promise for developing novel peptide-based therapeutics.