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High-throughput phosphotyrosine profiling using SH2 domains. Mol Cell 2007 Jun 22;26(6):899-915

Date

06/26/2007

Pubmed ID

17588523

DOI

10.1016/j.molcel.2007.05.031

Scopus ID

2-s2.0-34250641161 (requires institutional sign-in at Scopus site)   150 Citations

Abstract

Protein tyrosine phosphorylation controls many aspects of signaling in multicellular organisms. One of the major consequences of tyrosine phosphorylation is the creation of binding sites for proteins containing Src homology 2 (SH2) domains. To profile the global tyrosine phosphorylation state of the cell, we have developed proteomic binding assays encompassing nearly the full complement of human SH2 domains. Here we provide a global view of SH2 domain binding to cellular proteins based on large-scale far-western analyses. We also use reverse-phase protein arrays to generate comprehensive, quantitative SH2 binding profiles for phosphopeptides, recombinant proteins, and entire proteomes. As an example, we profiled the adhesion-dependent SH2 binding interactions in fibroblasts and identified specific focal adhesion complex proteins whose tyrosine phosphorylation and binding to SH2 domains are modulated by adhesion. These results demonstrate that high-throughput comprehensive SH2 profiling provides valuable mechanistic insights into tyrosine kinase signaling pathways.

Author List

Machida K, Thompson CM, Dierck K, Jablonowski K, Kärkkäinen S, Liu B, Zhang H, Nash PD, Newman DK, Nollau P, Pawson T, Renkema GH, Saksela K, Schiller MR, Shin DG, Mayer BJ

Authors

Debra K. Newman PhD Investigator in the Blood Research Institute department at BloodCenter of Wisconsin
Debra K. Newman PhD Professor in the Pharmacology and Toxicology department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Animals
Cell Adhesion
Fibroblasts
Focal Adhesions
Humans
Mice
Multiprotein Complexes
NIH 3T3 Cells
Peptides
Phosphorylation
Phosphotyrosine
Protein Array Analysis
Protein Processing, Post-Translational
Protein-Tyrosine Kinases
Proteome
Recombinant Proteins
Signal Transduction
src Homology Domains