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Cited by 516Chemical Synthesis and AnalysisProtein Structure and DynamicsEvolution and Genetic Dynamics

High-resolution mapping of protein sequence-function relationships

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Fowler, Douglas M, Araya, Carlos L, Fleishman, Sarel J, Kellogg, Elizabeth H, Stephany, Jason J, Baker, David, Fields, Stanley·2010·Nature Methods
DOI 10.1038/nmeth.1492PMC PMC2938879
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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.

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Research works printed but not linked: 29

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Authors
Fowler, D. M., Araya, C. L., Fleishman, S. J., Kellogg, E. H., Stephany, J. J., Baker, D., & Fields, S.
Affiliations
  • University of Washington (US)
  • Howard Hughes Medical Institute (US)
Corresponding author
Fields, Stanley
ORCID
Venue
Nature Methods
Publisher
Springer Science and Business Media LLC
Published
2010-08-15
Volume
7
Issue
9
Pages
741-746
ISSN
  • 1548-7091 (ISSN-L)
  • 1548-7105
DOI
10.1038/nmeth.1492
PMC
PMC2938879
OpenAlex
W2151887741
Cited by
516
References
1
Citations by year
2026: 27 · 2025: 31 · 2024: 42 · 2023: 35 · 2022: 38 · 2021: 28 · 2020: 29 · 2019: 40 · 2018: 39 · 2017: 25 · 2016: 35 · 2015: 43 · 2014: 34 · 2013: 31 · 2012: 28
FWCI
8.50
Type
journal article
Language
en
Open access
Licence
Locations
  • Nature Methods (publishedVersion)
  • PubMed (publishedVersion)
  • PubMed Central (acceptedVersion)
Funders
  • Howard Hughes Medical Institute
  • National Institute of General Medical Sciences (F32GM084699, P41GM103533)
  • National Center for Research Resources (P41RR011823)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Chemical Synthesis and Analysis
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Structure and Dynamics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Evolution and Genetic Dynamics
Keywords
phage display, WW domain, peptide ligand binding, protein variants, next-generation sequencing
MeSH terms
DNA, Humans, Proteins, Structure-Activity Relationship, Sequence Analysis, DNA, Peptide Library, Databases, Nucleic Acid, Protein Array Analysis, High-Throughput Screening Assays

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