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Nat Methods. 2010 Sep;7(9):741-6. doi: 10.1038/nmeth.1492. Epub 2010 Aug 15.

High-resolution mapping of protein sequence-function relationships.

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  • 1Department of Genome Sciences, University of Washington, Seattle, WA, USA.

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.

PMID:
20711194
[PubMed - indexed for MEDLINE]
PMCID:
PMC2938879
Free PMC Article

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