Analysis for Signaling Networks by Large-scale Monitoring of Phosphorylated Tyrosines

• Project/Area Number: 26830125
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Genome biology
• Research Institution: Kyoto University
• Principal Investigator: Sugiyama Naoyuki 京都大学, 薬学研究科(研究院), 准教授 (50545704)
• Project Period (FY): 2014-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥3,900,000 (Direct Cost: ¥3,000,000、Indirect Cost: ¥900,000); Fiscal Year 2015: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2014: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: プロテオーム / シグナル伝達 / リン酸化プロテオミクス / プロテオミクス / リン酸化プロテオーム

Outline of Final Research Achievements

In this study, a highly selective method for phosphotyrosine (pY)-containing peptides was developed to comprehensively identify pYs and analyze cellular signaling networks based on the data.
By a combination use of metal oxide chromatography and immunoprecipitation using anti-pY antibody or in vitro kinase reaction with recombinant tyrosine kinases, phosphotyrosines in biological samples were successfully identified in large scale. Using this approach, molecular-targeting agents of a cancer were evaluated and regulated phosphorylated sites were determined.

Research Products

• [Journal Article] Uncovering Phosphorylation-Based Specificities through Functional Interaction Networks. (2016)
  • Author(s): Wagih O, Sugiyama N, Ishihama Y, Beltrao P
  • Journal Title: Molecular & Cellular Proteomics Volume: 15 Issue: 1 Pages: 236-45
  • DOI: 10.1074/mcp.m115.052357
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Human mesenchymal stem cell-engineered hepatic cell sheets accelerate liver regeneration in mice (2015)
  • Author(s): Itaba N, Matsumi Y, Okinaka K, Ashla AA, Kono Y, Osaki M, Morimoto M, Sugiyama N, Ohashi K, Okano T, Shiota G
  • Journal Title: Scientific Reports Volume: 5 Issue: 1 Pages: 16169-16169
  • DOI: 10.1038/srep16169
  • NAID: 120005822792
  • Peer Reviewed / Open Access
• [Journal Article] Extended coverage of singly and multiply phosphorylated peptides from a single titanium dioxide microcolumn (2015)
  • Author(s): Masaki Wakabayashi, Yutaka Kyono, Naoyuki Sugiyama, Yasushi Ishihama
  • Journal Title: Analytical chemistry Volume: 87 Issue: 20 Pages: 10213-21
  • DOI: 10.1021/acs.analchem.5b01216
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] Systematic profiling of the bacterial phosphoproteome reveals bacterium-specific features of phosphorylation. (2015)
  • Author(s): Lin MH, Sugiyama N, Ishihama Y.
  • Journal Title: Sci Signal. Volume: 8 Issue: 394
  • DOI: 10.1126/scisignal.aaa3117
  • NAID: 120005682544
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] LATS1 and LATS2 phosphorylate CDC26 to modulate assembly of the tetratricopeptide repeat subcomplex of APC/C. (2015)
  • Author(s): Masuda K, Chiyoda T, Sugiyama N, Segura-Cabrera A, Kabe Y, Ueki A, Banno K, Suematsu M, Aoki D, Ishihama Y, Saya H, Kuninaka S
  • Journal Title: PLoS One Volume: 10 Issue: 2 Pages: 1-22
  • DOI: 10.1371/journal.pone.0118662
  • Peer Reviewed / Open Access
• [Journal Article] PCTK1 Regulates Integrin-Dependent Spindle Orientation via Protein Kinase A Regulatory Subunit KAP0 and Myosin X. (2015)
  • Author(s): Iwano S, Satou A, Matsumura S, Sugiyama N, Ishihama Y, Toyoshima F.
  • Journal Title: Mol Cell Biol Volume: 35 Issue: 7 Pages: 1197-208
  • DOI: 10.1128/mcb.01017-14
  • Peer Reviewed
• [Journal Article] Large-Scale Identification of Phosphorylation Sites for Profiling Protein Kinase Selectivity. (2014)
  • Author(s): Imamura H, Sugiyama N, Wakabayashi M, Ishihama Y.
  • Journal Title: J Proteome Res. Volume: 13 Issue: 7 Pages: 3410-19
  • DOI: 10.1021/pr500319y
  • Peer Reviewed / Acknowledgement Compliant
• [Presentation] 大規模プロテオミクスのための解析ワークフローと統合データベース （jPOST） (2015)
  • Author(s): 杉山直幸
  • Organizer: 生命医薬情報学連合大会 2015 年大会
  • Place of Presentation: 京都大学宇治キャンパスおうばくプラザ（京都・宇治）
  • Year and Date: 2015-10-29
  • Invited
• [Presentation] Evaluation of Protein Kinase Inhibitors Using Motif-Targeting Phosphoproteomics (2015)
  • Author(s): Naoyuki Sugiyama, Kosuke Ogata, Tatsuya Yazaki, Yuu Hayashi, Shunsuke Takagi, Masaki Wakabayashi, Yasushi Ishihama
  • Organizer: 14th Human Proteome Organization World Congress (HUPO2015)
  • Place of Presentation: Vancouver (Canada)
  • Year and Date: 2015-09-27
• [Presentation] モチーフ標的定量法を用いた分子標的薬のリン酸化プロテオームプロファイリング (2015)
  • Author(s): 小形公亮、矢崎達也、若林真樹、杉山直幸、石濱泰
  • Organizer: 日本プロテオーム学会2015年会
  • Place of Presentation: くまもと森都心プラザ（熊本・熊本）
  • Year and Date: 2015-07-23
• [Presentation] 高選択的基質ペプチドを用いたヒトキノーム活性測定 (2015)
  • Author(s): 杉山直幸、坂本 大、今村春菜、Pasrawin Taechawattananant、若林真樹、石濱 泰
  • Organizer: 日本プロテオーム学会2015年会
  • Place of Presentation: くまもと森都心プラザ（熊本・熊本）
  • Year and Date: 2015-07-23
• [Presentation] Monitoring Kinome Activities Using the Highly Specific and Sensitive Substrate Peptides (2014)
  • Author(s): Naoyuki Sugiyama, Haruna Imamura, Pasrawin Taechawattananant, Dai Sakamoto, Masaki Wakabayashi, Yasushi Ishihama
  • Organizer: 13th Human Proteome Organization World Congress
  • Place of Presentation: Madrid
  • Year and Date: 2014-10-06
• [Presentation] プロテオミクスのデータ処理法とツール (2014)
  • Author(s): 杉山直幸
  • Organizer: 日本プロテオーム学会2014年会
  • Place of Presentation: つくば
  • Year and Date: 2014-07-17
• [Remarks] 京都大学薬学研究科 石濱研究室
  • URL: http://www.pharm.kyoto-u.ac.jp/seizai/