Development of molecular diagnosis system of cancer drug resistance using absolute quantitative proteomics

• Project/Area Number: 23701084
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Tumor diagnosis
• Research Institution: Ehime University
• Principal Investigator: TAKEMORI Nobuaki 愛媛大学, プロテオ医学研究センター, 講師 (40533047)
• Project Period (FY): 2011 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2012: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2011: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
• Keywords: 薬剤耐性 / 絶対定量プロテオミクス / 定量MRM法 / 小麦無細胞タンパク質合成系 / コムギ無細胞タンパク質合成系

Research Abstract

We have established a molecular diagnosis system for quantifying proteomic alterations of cancer drug resistance using wheat germ cell-free protein synthesis system and quantitative mass spectrometry. Stable isotope (SI) labeled recombinant proteins as an internal standard for protein quantification were in vitro synthesized using wheat cell-free synthesis system. We selected a set of digested peptides for each protein based on their ionization efficiency and established multiple reaction monitoring (MRM) assay using triple quadrupole mass spectrometer. We are currently constructing an absolute quantitative platform of human biopsy samples by combining MRM assay and synthesized SI-labeled protein library, which allows an accurate measurement of the absolute amount of targeted proteins from a limited amount of tissue samples.

Research Products

• [Journal Article] Adseverin: a novel cisplatin-resistant marker in the human bladder cancer cell line HT1376 identified by quantitative proteomic analysis (2012)
  • Author(s): Miura N, Takemori N, Kikugawa T, Tanji N, Higashiyama S, Yokoyama M
  • Journal Title: Mol Oncol. Volume: 6 Pages: 311-322
  • Peer Reviewed
• [Journal Article] Adseverin: a novel cisplatin-resistant marker in the human bladder cancer cell line HT1376 identified by quantitative proteomic analysis. (2012)
  • Author(s): Miura N, Takemori N, Kikugawa T, Tanji N, Higashiyama S, Yokoyama M.
  • Journal Title: Molecular Oncology Volume: 6 Issue: 3 Pages: 311-322
  • DOI: 10.1016/j.molonc.2011.12.002
  • Peer Reviewed
• [Presentation] Absolute quantitative analysis of urinary proteome alterations using wheat germ cell free protein synthesis and MRM mass spectrometry (2013)
  • Author(s): Takemori N, Takemori A, Higashiyama S
  • Organizer: 11th HUPO World Congress
  • Place of Presentation: Hynes Convention Center, Boston, U.S.A
  • Year and Date: 2013-09-13
• [Presentation] SIRPLDB:安定同位体標識リコンビナントタンパク質ライブラリーの定量プロテオーム解析への活用に向けた情報データベースの構築 (2012)
  • Author(s): 武森文子、武森信暁、東山繁樹
  • Organizer: 第85回日本生化学会大会
  • Place of Presentation: マリンメッセ福岡、福岡
  • Year and Date: 2012-12-16
• [Presentation] A high-throughput targeted approach for the absolute quantification of human and mouse proteome alterations (2012)
  • Author(s): 武森信暁、武森文子、青島正人、田中ゆき、東 山 繁 樹
  • Organizer: 第85回日本生化学会大会
  • Place of Presentation: マリンメッセ福岡、福岡
  • Year and Date: 2012-12-16
• [Presentation] Accelerating quantitative proteomics research with wheat germ cell-free protein synthesis system. (2012)
  • Author(s): Takemori N
  • Organizer: Protein Island Matsuyama International Symposium 2012
  • Place of Presentation: 松山全日空ホテル、愛媛
  • Year and Date: 2012-09-25
• [Presentation] SIRPL: Construction of a stable isotope-labeled recombinant protein library for the absolute quantitative analysis of mouse membrane proteome (2012)
  • Author(s): Takemori N, Takemori A, Matsuoka K, Matsushita N, Takeda H, Sawasaki T, Endo Y, Higashiyama S
  • Organizer: The 60th American Society for Mass Spectrometry (ASMS) Conference
  • Place of Presentation: Vancouver Convention Center, Vancouver, Canada
  • Year and Date: 2012-05-24
• [Presentation] 安定同位体標識リコンビナントタンパク質ライブラリーを用いたマウストランスメンブレンプロテオームの絶対定量測定 (2011)
  • Author(s): 武森信暁、武森文子、松岡和弘、松下夏樹、澤崎達也、遠藤弥重太、東山繁樹
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 国立京都国際会館、京都
  • Year and Date: 2011-09-23
• [Presentation] An absolute quantification method of mouse transmembrane proteome using a combination of wheat cell-free synthesis and MRM mass spectrometry (2011)
  • Author(s): Takemori N, Takemori A, Matsuoka K, Matsushita N, Sawasaki T, Endo Y, Higashiyama S
  • Organizer: 10th HUPO World Congress
  • Place of Presentation: PALEXPO, Geneva, Switzerland
  • Year and Date: 2011-09-06
• [Presentation] An absolute quantification method of mouse transmembrane proteome using a combination of wheat cell-free synthesis and MRM mass spectrometry (2011)
  • Author(s): Takemori N, Takemori A, Matsuoka K, Matsushita N, Sawasaki T, Endo Y, and Higashiyama S
  • Organizer: 10th HUPO World Congress
  • Place of Presentation: PALEXPO Geneva, Switzerland
• [Presentation] SIRPL: Construction of a stable isotope-labeled recombinant protein library for the absolute quantitative analysis of mouse membrane proteome.
  • Author(s): Takemori N, Takemori A, Matsuoka K, Matsushita N, Takeda H, Sawasaki T, Endo Y, Higashiyama S
  • Organizer: The 60th American Society for Mass Spectrometry (ASMS) Conference
  • Place of Presentation: Vancouver Convention Center, Vancouver, Canada
• [Presentation] Absolute quantitative analysis of urinary proteome alterations using wheat germ cell free protein synthesis and MRM mass spectrometry.
  • Author(s): Takemori N, Takemori A, Higashiyama S
  • Organizer: 11th HUPO World Congress
  • Place of Presentation: Hynes Convention Center, Boston, U.S.A
• [Presentation] SIRPLDB：安定同位体標識リコンビナントタンパク質ライブラリーの定量プロテオーム解析への活用に向けた情報データベースの構築
  • Author(s): 武森文子、武森信暁、東山繁樹
  • Organizer: 第85回日本生化学会大会
  • Place of Presentation: マリンメッセ福岡、福岡
• [Presentation] A high-throughput targeted approach for the absolute quantification of human and mouse proteome alterations.
  • Author(s): 武森信暁、武森文子、青島正人、田中ゆき、東山繁樹
  • Organizer: 第85回 日本生化学会大会
  • Place of Presentation: マリンメッセ福岡、福岡