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ファイル | 記述 | サイズ | フォーマット | |
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j.crmeth.2021.100138.pdf | 2.35 MB | Adobe PDF | 見る/開く |
タイトル: | Motif-centric phosphoproteomics to target kinase-mediated signaling pathways |
著者: | Tsai, Chia-Feng Ogata, Kosuke https://orcid.org/0000-0002-0634-3990 (unconfirmed) Sugiyama, Naoyuki Ishihama, Yasushi https://orcid.org/0000-0001-7714-203X (unconfirmed) |
著者名の別形: | 小形, 公亮 杉山, 直幸 石濱, 泰 |
キーワード: | motif-centric phosphoproteome in vitro kinase reaction phosphopeptide enrichment isobaric tag boosting MS signal tyrosine phosphoproteome TMT quantitation EGFR signaling network kinase-substrate relationship |
発行日: | Jan-2022 |
出版者: | Elsevier BV |
誌名: | Cell Reports Methods |
巻: | 2 |
号: | 1 |
論文番号: | 100138 |
抄録: | Identifying cellular phosphorylation pathways based on kinase-substrate relationships is a critical step to understanding the regulation of physiological functions in cells. Mass spectrometry-based phosphoproteomics workflows have made it possible to comprehensively collect information on individual phosphorylation sites in a variety of samples. However, there is still no generic approach to uncover phosphorylation networks based on kinase-substrate relationships in rare cell populations. Here, we describe a motif-centric phosphoproteomics approach combined with multiplexed isobaric labeling, in which in vitro kinase reactions are used to generate targeted phosphopeptides, which are spiked into one of the isobaric channels to increase detectability. Proof-of-concept experiments demonstrate selective and comprehensive quantification of targeted phosphopeptides by using multiple kinases for motif-centric channels. More than 7, 000 tyrosine phosphorylation sites were quantified from several tens of micrograms of starting materials. This approach enables the quantification of multiple phosphorylation pathways under physiological or pathological regulation in a motif-centric manner. |
記述: | 細胞内リン酸化修飾の大規模計測に成功 --極微量試料からのリン酸化経路解析も可能に--. 京都大学プレスリリース. 2022-02-01. |
著作権等: | © 2021 The Author(s). This is an open access article under the Creative Commons Attribution 4.0 International license. |
URI: | http://hdl.handle.net/2433/267848 |
DOI(出版社版): | 10.1016/j.crmeth.2021.100138 |
PubMed ID: | 35474870 |
関連リンク: | https://www.kyoto-u.ac.jp/ja/research-news/2022-02-01 |
出現コレクション: | 学術雑誌掲載論文等 |
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