High-throughput phosphotyrosine profiling using SH2 domains |
| |
Authors: | Machida Kazuya Thompson Christopher M Dierck Kevin Jablonowski Karl Kärkkäinen Satu Liu Bernard Zhang Haimin Nash Piers D Newman Debra K Nollau Peter Pawson Tony Renkema G Herma Saksela Kalle Schiller Martin R Shin Dong-Guk Mayer Bruce J |
| |
Institution: | Raymond and Beverly Sackler Laboratory of Genetics and Molecular Medicine, Department of Genetics and Developmental Biology, University of Connecticut Health Center, Farmington, CT 06030, USA. |
| |
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. |
| |
Keywords: | |
本文献已被 PubMed 等数据库收录! |
|