High resolution mapping of protein sequence-function relationships in Human Yap65 CA0002
High resolution mapping of protein sequence-function relationships in Human Yap65 CA0002
The goal of this study was to demonstrate the power of high-throughput sequencing technologies in tracking the effects of mutations on fitness.
We present a large-scale approach to investigate the functional consequences of sequence variation in a protein.  The approach entails the display of hundreds of thousands of protein variants, moderate selection for activity, and high throughput DNA sequencing to quantify the performance of each variant.  Using this strategy, we tracked  the performance of >600,000 variants of a human WW domain after three and six rounds of selection by phage display for binding to its peptide ligand.  Binding properties of these variants defined a high-resolution map of mutational preference across the WW domain; each position possessed unique features that could not be captured by a few representative mutations.  Our approach could be applied to many in vitro or in vivo protein assays, providing a general means for understanding how protein function relates to sequence.