High-resolution mapping of protein sequence-function relationships


Abstract

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 had 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.

Submission Details

ID: Qhd2UUxv3

Submitter: Connie Wang

Submission Date: Oct. 24, 2016, 4:27 p.m.

Version: 1

Publication Details
Fowler DM;Araya CL;Fleishman SJ;Kellogg EH;Stephany JJ;Baker D;Fields S,Nat Methods (2010) High-resolution mapping of protein sequence-function relationships. PMID:20711194
Additional Information

Sequence Assay Result Units