Abstract
In this chapter, we discuss a method to determine the affinity and specificity of nearly all single-point mutants for a full-length protein binder. This method combines deep sequencing, comprehensive mutagenesis, yeast surface display, and fluorescence-activated cell sorting. This approach has been used to study sequence-function relationships for protein-protein interactions. The data can be used to determine the fine conformational epitope on the protein binder.
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Acknowledgments
This work was supported by NSF CAREER (Award #1254238) to T.A.W. and a NIH T32 Biotechnology Training Grant (Award # T32-GM110523) to A.M.C.
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Medina-Cucurella, A.V., Whitehead, T.A. (2018). Characterizing Protein-Protein Interactions Using Deep Sequencing Coupled to Yeast Surface Display. In: Marsh, J. (eds) Protein Complex Assembly. Methods in Molecular Biology, vol 1764. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7759-8_7
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DOI: https://doi.org/10.1007/978-1-4939-7759-8_7
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