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Characterizing Protein-Protein Interactions Using Deep Sequencing Coupled to Yeast Surface Display

  • Angelica V. Medina-Cucurella
  • Timothy A. Whitehead
Part of the Methods in Molecular Biology book series (MIMB, volume 1764)

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.

Key words

Deep sequencing Yeast surface display Nicking mutagenesis FACS Conformational epitope mapping 

Notes

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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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  • Angelica V. Medina-Cucurella
    • 1
  • Timothy A. Whitehead
    • 1
    • 2
  1. 1.Department of Chemical Engineering and Materials ScienceMichigan State UniversityEast LansingUSA
  2. 2.Department of Biosystems and Agricultural EngineeringMichigan State UniversityEast LansingUSA

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