title: Epitope-Specific Binder Design by Yeast Surface Display.
abstract: Yeast surface display is commonly used to engineer affinity and design novel molecular interaction. By alternating positive and negative selections, yeast display can be used to engineer binders that specifically interact with the target protein at a defined site. Epitope-specific binders can be useful as inhibitors if they bind the target molecule at functionally important sites. Therefore, an efficient method of engineering epitope specificity should help with the engineering of inhibitors. We describe the use of yeast surface display to design single domain monobodies that bind and inhibit the activity of the kinase Erk-2 by targeting a conserved surface patch involved in protein-protein interaction. The designed binders can be used to disrupt signaling in the cell and investigate Erk-2 function in vivo. The described protocol is general and can be used to design epitope-specific binders of an arbitrary protein.
authors.affiliation: Department of Chemical and Biological Engineering, University at Buffalo, 905 Furnas Hall, Buffalo, NY, 14260, USA.
authors.forename: Jasdeep K
authors.initials: JK
authors.lastname: Mann
authors.forename: Sheldon
authors.initials: S
authors.lastname: Park
mesh_terms: D001665:Binding Sites / Q000187:drug effects; D000939:Epitopes / Q000235:genetics*; D006801:Humans; D019950:Mitogen-Activated Protein Kinase 1 / Q000037:antagonists & inhibitors / Q000737:chemistry*; D009154:Mutation; D019151:Peptide Library; D011485:Protein Binding; D015202:Protein Engineering; D047428:Protein Kinase Inhibitors / Q000378:metabolism / Q000494:pharmacology; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism; D061905:Single-Domain Antibodies / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology
chemical_list: D000939:Epitopes; D019151:Peptide Library; D047428:Protein Kinase Inhibitors; D061905:Single-Domain Antibodies; C535150:MAPK1 protein, human; D019950:Mitogen-Activated Protein Kinase 1