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gtc.13063.pdf | 2.26 MB | Adobe PDF | 見る/開く |
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dc.contributor.author | Niinae, Tomoya | en |
dc.contributor.author | Sugiyama, Naoyuki | en |
dc.contributor.author | Ishihama, Yasushi | en |
dc.contributor.alternative | 新苗, 智也 | ja |
dc.contributor.alternative | 杉山, 直幸 | ja |
dc.contributor.alternative | 石濱, 泰 | ja |
dc.date.accessioned | 2024-03-06T06:09:49Z | - |
dc.date.available | 2024-03-06T06:09:49Z | - |
dc.date.issued | 2023-10 | - |
dc.identifier.uri | http://hdl.handle.net/2433/287218 | - |
dc.description.abstract | Three representative protein kinases with different substrate preferences, ERK1 (Pro-directed), CK2 (acidophilic), and PKA (basophilic), were used to investigate phosphorylation sequence motifs in substrate pools consisting of the proteomes from three different cell lines, MCF7 (human mammary carcinoma), HeLa (human cervical carcinoma), and Jurkat (human acute T-cell leukemia). Specifically, recombinant kinases were added to the cell-extracted proteomes to phosphorylate the substrates in vitro. After trypsin digestion, the phosphopeptides were enriched and subjected to nanoLC/MS/MS analysis to identify their phosphorylation sites on a large scale. By analyzing the obtained phosphorylation sites and their surrounding sequences, phosphorylation motifs were extracted for each kinase-substrate proteome pair. We found that each kinase exhibited the same set of phosphorylation motifs, independently of the substrate pool proteome. Furthermore, the identified motifs were also consistent with those found using a completely randomized peptide library. These results indicate that cell-extracted proteomes can provide kinase phosphorylation motifs with sufficient accuracy, even though their sequences are not completely random, supporting the robustness of phosphorylation motif identification based on phosphoproteome analysis of cell extracts as a substrate pool for a kinase of interest. | en |
dc.language.iso | eng | - |
dc.publisher | Molecular Biology Society of Japan | en |
dc.publisher | Wiley | en |
dc.rights | © 2023 The Authors. Genes to Cells published by Molecular Biology Society of Japan and John Wiley & Sons Australia, Ltd. | en |
dc.rights | This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. | en |
dc.rights.uri | http://creativecommons.org/licenses/by/4.0/ | - |
dc.subject | Kinase substrate | en |
dc.subject | Phosphoproteomics | en |
dc.subject | Phosphorylation motif | en |
dc.subject | Protein kinase | en |
dc.title | Validity of the cell‐extracted proteome as a substrate pool for exploring phosphorylation motifs of kinases | en |
dc.type | journal article | - |
dc.type.niitype | Journal Article | - |
dc.identifier.jtitle | Genes to Cells | en |
dc.identifier.volume | 28 | - |
dc.identifier.issue | 10 | - |
dc.identifier.spage | 727 | - |
dc.identifier.epage | 735 | - |
dc.relation.doi | 10.1111/gtc.13063 | - |
dc.textversion | publisher | - |
dc.identifier.pmid | 37658684 | - |
dcterms.accessRights | open access | - |
datacite.awardNumber | 17H03605 | - |
datacite.awardNumber | 18H04799 | - |
datacite.awardNumber | 20H04845 | - |
datacite.awardNumber | 21H02459 | - |
datacite.awardNumber | 21H02466 | - |
datacite.awardNumber | 21J15068 | - |
datacite.awardNumber | 23H04924 | - |
datacite.awardNumber.uri | https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-17H03605/ | - |
datacite.awardNumber.uri | https://kaken.nii.ac.jp/grant/KAKENHI-PUBLICLY-18H04799/ | - |
datacite.awardNumber.uri | https://kaken.nii.ac.jp/grant/KAKENHI-PUBLICLY-20H04845/ | - |
datacite.awardNumber.uri | https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21H02459/ | - |
datacite.awardNumber.uri | https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21H02466/ | - |
datacite.awardNumber.uri | https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-21J15068/ | - |
datacite.awardNumber.uri | https://kaken.nii.ac.jp/grant/KAKENHI-PLANNED-23H04924/ | - |
dc.identifier.pissn | 1356-9597 | - |
dc.identifier.eissn | 1365-2443 | - |
jpcoar.funderName | 日本学術振興会 | ja |
jpcoar.funderName | 日本学術振興会 | ja |
jpcoar.funderName | 日本学術振興会 | ja |
jpcoar.funderName | 日本学術振興会 | ja |
jpcoar.funderName | 日本学術振興会 | ja |
jpcoar.funderName | 日本学術振興会 | ja |
jpcoar.funderName | 日本学術振興会 | ja |
jpcoar.awardTitle | キノームの生理的基質同定に基づく細胞内シグナルパスウェイ大規模解析 | ja |
jpcoar.awardTitle | キノームの機能制御メカニズムの解明 | ja |
jpcoar.awardTitle | 活性制御部位関連ペプチドによるキノーム活性測定法の開発 | ja |
jpcoar.awardTitle | 大規模プロテオフォーム解析法の開発とその全体像の理解 | ja |
jpcoar.awardTitle | ヒトキノームの基質認識モチーフ探索 | ja |
jpcoar.awardTitle | キナーゼ基質網羅的同定とゲノム変異依存的リン酸化シグナル解析 | ja |
jpcoar.awardTitle | タンパク質寿命の大規模計測技術と寿命制御原理 | ja |
出現コレクション: | 学術雑誌掲載論文等 |
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