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Updated: Jun 12, 2025
05:49
Protein Engineering by Yeast Surface Display
Published on: November 29, 2024
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Protocol for engineering binding domains to recognize ligand-bound receptors by using yeast surface display.
Markus Dobersberger1, Delia Sumesgutner2, Charlotte U Zajc2
1Department of Chemistry, Institute of Biochemistry, BOKU University, Vienna 1190, Austria.
STAR Protocols
|September 25, 2024
Summary
This study introduces a yeast surface display method to find proteins that bind to receptors only when they are bound to their ligands. This protein engineering technique enhances the discovery of specific molecular interactions.
Area of Science:
- Protein engineering
- Molecular biology
- Biotechnology
Background:
- Yeast surface display is a powerful protein engineering tool.
- Selecting binders for ligand-bound receptors requires specific methods.
- Existing techniques may not efficiently isolate these specific binders.
Purpose of the Study:
- To present a detailed protocol for yeast surface display to enrich binders targeting ligand-bound receptors.
- To demonstrate the applicability of this method for protein engineering and binder discovery.
- To provide a framework adaptable to various binder scaffolds and ligand-receptor systems.
Main Methods:
- Utilizing yeast surface display for protein library construction.
- Employing magnetic bead selection and fluorescence-activated cell sorting (FACS) for enrichment.
- Incorporating random mutagenesis for library diversification.
- Performing library sequencing and bioinformatics analysis for clone characterization.
Main Results:
- Successful enrichment of binders that specifically recognize ligand-bound receptors.
- Demonstration of the protocol using rcSso7d binders and epidermal growth factor (EGF)-epidermal growth factor receptor (EGFR) complexes.
- Identification and analysis of enriched protein clones.
Conclusions:
- The presented yeast surface display protocol is effective for isolating binders of ligand-bound receptors.
- This method offers a versatile platform for protein engineering and drug discovery.
- The protocol can be adapted for diverse biological systems and binder scaffolds.
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