High-resolution mapping of protein sequence-function relationships

TitleHigh-resolution mapping of protein sequence-function relationships
Publication TypeJournal Article
Year of Publication2010
AuthorsFowler, D. M., Araya C. L., Fleishman S. J., Kellogg E. H., Stephany J. J., Baker D., & Fields S.
JournalNature methods
Volume7
Issue9
Pagination741-6
Date Published2010 Sep
ISSN1548-7105
KeywordsCollaborative Publication, Databases, Nucleic Acid, DNA, High-Throughput Screening Assays, Humans, Peptide Library, Protein Array Analysis, Proteins, Sequence Analysis, DNA, Structure-Activity Relationship
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.

Alternate JournalNat. Methods
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