オーラルセッション
- 第2日 5月16日（水） 10:25～10:45 C会場（星雲2）
-
2C-O1-P-1025 PDF
網羅的チロシンリン酸化プロテオミクス解析法の構築
The phosphosignaling acts as regulator of cell proliferation, differentiation, environmental response. The failure of phosphosignaling, especially phosphotyrosine signaling, leads to oncogenesis. Thus, tyrosine kinase family is an important target in cancer therapy. Comprehensive phosphotyrosine proteomics would greatly contribute the discovery of novel cancer therapeutic targets and the establishment of efficient therapeutic strategies.
When performing large-scale phosphoproteomics using the IMAC, MOAC, and HAMMOC method, the ratio of phosphorylated tyrosine sites in the identified phosphorylated peptides is usually limited less than 1%. In order to improve the identification number of phosphotyrosine sites, previous study performed immunoprecipitation with antibody against phosphotyrosine site. In this study, we tried to improve the identification number of phosphorylated tyrosine by examining several conditions during immunoprecipitation and LC-MSMS analysis.
As a result, we succeeded in identifying more than 1,000 phosphotyrosine sites per sample. Also, we measured modulation of phosphotyrosine signaling in cells with or without treatment of tyrosine kinase inhibitor. Furthermore, kinome analysis was performed by combining phospho-proteomics method by Fe-IMAC with optimized tyrosine phosphorylation proteomics method.
Our phosphotyrosine proteomics is expected to lead to the identification of novel therapeutic targets and diagnostic markers for drug sensitivity.