Fig. 1.

A streamlined process required for PPI characterization using deep sequencing and mutagenesis analysis. (a-c) Requirements for the pipeline: the binding activity of two proteins is measured using yeast surface display coupled to flow cytometry, and the relative dissociation constant is determined using yeast clonal titrations. The top panel is adapted from Chao et al. [9]. (d) The workflow covered in this chapter to characterize protein-protein interactions. (e, f) Deep sequencing results can be visualized as a heatmap and used to determine the conformational epitope of one member of the interaction