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: July 31, 2017, 11:46 a.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:20711194Additional Information
Study Summary
| Number of data points | 416 |
| Proteins | WW domain hYAP65 |
| Unique complexes | 397 |
| Assays/Quantities/Protocols | Experimental Assay: Phage display binding and then sequencing abundance |
| Libraries | Allowed mutations in 25 residue region due to illumina restraints, only looking at single mutants here |
Structure view and single mutant data analysis
Study data
| Colors: | D | E | R | H | K | S | T | N | Q | A | V | I | L | M | F | Y | W | C | G | P |
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Data Distribution
Studies with similar sequences (approximate matches)