Stepwise Evolution Improves Identification of Diverse Peptides Binding to a Protein Target

Area of Science:

• Biochemistry and Molecular Biology
• Proteomics and Peptidomics
• Drug Discovery and Development

Background:

• Developing selective peptide binders for target proteins is crucial for research and therapeutics.
• Existing methods for peptide discovery can be limited in scope and efficiency.
• Streptavidin-biotin interactions are well-studied but novel peptide mimics are of interest.

Purpose of the Study:

• To present a novel, reproducible, and digitally controlled method for discovering and evolving peptide binders.
• To demonstrate the application of this method for identifying high-affinity and high-specificity peptide hotspots.
• To discover novel peptide binders to streptavidin that mimic biotin interactions.

Main Methods:

• Combined rational peptide array library design with parallel screening and stepwise evolution.
• Applied the method to synthesize diverse peptides, including linear, cyclic, and those with non-natural amino acids.
• Used streptavidin as a model protein target for evolving peptide binders.

Main Results:

• Successfully discovered novel peptide hotspots with potential for high-affinity binding.
• Demonstrated systematic evolution of these hotspots into high-specificity binding peptides.
• Identified novel peptides that mimic key biotin-streptavidin binding interactions.

Conclusions:

• The developed method offers a powerful and versatile platform for peptide binder discovery and optimization.
• This approach enables exploration of a diverse chemical space for peptide synthesis.
• The novel streptavidin binders represent a significant advancement in understanding protein-peptide interactions.