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タイトル: Large-scale determination of absolute phosphorylation stoichiometries in human cells by motif-targeting quantitative proteomics.
著者: Tsai, Chia-Feng
Wang, Yi-Ting
Yen, Hsin-Yung
Tsou, Chih-Chiang
Ku, Wei-Chi
Lin, Pei-Yi
Chen, Hsuan-Yu
Nesvizhskii, Alexey I
Ishihama, Yasushi  kyouindb  KAKEN_id
Chen, Yu-Ju
著者名の別形: 石濱, 泰
キーワード: Biological sciences
Biochemistry
Systems biology
発行日: 27-Mar-2015
出版者: Nature Publishing Group
誌名: Nature communications
巻: 6
論文番号: 6622
抄録: 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-of-concept 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 >1,000 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.
著作権等: This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
URI: http://hdl.handle.net/2433/198827
DOI(出版社版): 10.1038/ncomms7622
PubMed ID: 25814448
出現コレクション:学術雑誌掲載論文等

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