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Encoding the Human Phosphoproteome in an Engineered Bacterial System
Author(s) -
Barber Karl,
Rinehart Jesse
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.614.33
Subject(s) - phosphoserine , phosphorylation , human proteome project , serine , proteome , computational biology , protein phosphorylation , biology , signal transduction , biochemistry , chemistry , proteomics , microbiology and biotechnology , protein kinase a , gene
Phosphorylation can potently alter protein folding and function, such as by modulating enzymatic activity, exposing scaffolding domains, or directly recruiting interaction partners. Although serine phosphorylation is the most common post‐translational modification in eukaryotes, the significance of the vast majority of phosphorylation sites across the human proteome remains unknown. Using an orthogonal translation system for phosphoserine (SepOTS), we are able to co‐translationally incorporate phosphoserine as a non‐standard amino acid in E. coli. We demonstrate the ability to synthesize and observe phosphoproteome‐scale peptide libraries representing tens of thousands of human phosphoproteins in a single experiment. We then deploy this massive‐scale peptide library to identify physiologically‐relevant phosphobinding protein interaction partners. Our synthetic human phosphoproteome is uniquely poised to answer lingering questions regarding the functional role of phosphorylation in signal transduction pathways by simultaneously monitoring thousands of human phosphosites. Support or Funding Information K.W.B. is supported by the National Science Foundation Graduate Research Fellowship under Grant No. DGE‐1122492. J.R. is supported by NIH R01 GM117230.