2015年3月
Large-scale determination of absolute phosphorylation stoichiometries in human cells by motif-targeting quantitative proteomics
NATURE COMMUNICATIONS
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- 巻
- 6
- 号
- 開始ページ
- 6622
- 終了ページ
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1038/ncomms7622
- 出版者・発行元
- NATURE PUBLISHING GROUP
Our ability to model the dynamics of signal transduction networks will depend on accurate methods to quantify levels of protein phosphorylation on a global scale. Here we describe a motif-targeting quantitation method for phosphorylation stoichiometry typing. Proteome-wide phosphorylation stoichiometry can be obtained by a simple phosphoproteomic workflow integrating dephosphorylation and isotope tagging with enzymatic kinase reaction. Proof-ofconcept experiments using CK2-, MAPK-and EGFR-targeting assays in lung cancer cells demonstrate the advantage of kinase-targeted complexity reduction, resulting in deeper phosphoproteome quantification. We measure the phosphorylation stoichiometry of 41,000 phosphorylation sites including 366 low-abundance tyrosine phosphorylation sites, with high reproducibility and using small sample sizes. Comparing drug-resistant and sensitive lung cancer cells, we reveal that post-translational phosphorylation changes are significantly more dramatic than those at the protein and messenger RNA levels, and suggest potential drug targets within the kinase-substrate network associated with acquired drug resistance.
- リンク情報
- ID情報
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- DOI : 10.1038/ncomms7622
- ISSN : 2041-1723
- PubMed ID : 25814448
- Web of Science ID : WOS:000353041300003