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Analysis of protein-protein interactions in human plasma using non-denaturing two-dimensional electrophoresis and mass spectrometry

Research Project

Project/Area Number 16550077
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Analytical chemistry
Research InstitutionEhime University

Principal Investigator

MANABE Takashi  Ehime University, Faculty of Science, professor, 理学部, 教授 (60094281)

Co-Investigator(Kenkyū-buntansha) SHIMAZAKI Yoji  Ehime University, Faculty of Science, instructor, 理学部, 助手 (80284389)
Project Period (FY) 2004 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2005: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2004: ¥2,400,000 (Direct Cost: ¥2,400,000)
Keywordshuman plasma / plasma proteins / non-denaturing / two-dimensional gel electrophoresis / micro gel / mass spectrometry / MALDI-TOF MS / protein interaction / マイクロ / ESI-MS / MS / 生理的条件
Research Abstract

Protein-protein interactions between human plasma proteins have been detected by micro two-dimensional electrophoresis (2-DE), comparing the patterns obtained under non-denaturing conditions and those obtained under partially denaturing conditions. The protein spots on the 2-DE gels, 157 spots on a non-denaturing micro 2-DE gel and 175 spots on a partially denaturing micro 2-DE gel, were subjected to structural analysis by mass spectrometry. For mass spectrometric analysis, matrix assisted laser desorption-time of flight mass spectrometry (MALDI-TOF MS) and database search (peptide mass fingerprinting, PMF) were employed. Integrating the information obtained, it was ascertained that structural information on human plasma proteins, including their status of complex-formation, can be analyzed. Although for the moment most of the protein researchers are using 2-DE under denaturing conditions, present study revealed that 2-DE under non-denaturing conditions will become more important by the advance of mass spectrometric methods in protein analysis.

Report

(3 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • Research Products

    (32 results)

All 2005 2004

All Journal Article (26 results) Book (6 results)

  • [Journal Article] Rapid Separation of Microorganism by Quartz Microchip Capillary Electrophoresis2005

    • Author(s)
      新谷智吉, 島村政基, 佐藤浩昭, 田尾博明, 真鍋 敬
    • Journal Title

      Anal. Sci. 21

      Pages: 57-60

    • NAID

      10014095894

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Alkaline Cleavage of Covalent Bonds in Chicken Insulin and Bovine alpha-Lactalbumin Analyzed by Matrix Assisted Laser Desorption/Ionization Mass Spectrometry2005

    • Author(s)
      真鍋 計, 金 亜
    • Journal Title

      Electrophoresis 26

      Pages: 257-267

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] High-efficiency protein extraction from polyacrylamide gels for molecular mass measurement by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry2005

    • Author(s)
      金 亜, 真鍋 敬
    • Journal Title

      Electrophoresis 26

      Pages: 1019-1028

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Direct targeting of human plasma for matrix-assisted laser desorption/ionization and analysis of plasma proteins by time of flight-mass spectrometry2005

    • Author(s)
      金 亜, 真鍋 敬
    • Journal Title

      Electrophoresis 26

      Pages: 2823-2834

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Detection of activity and mass spectrometric identification of mouse liver caboxylesterase and aldehyde dehydrogenase separated by non-denaturing two-dimensional electrophoresis after extraction with detergents2005

    • Author(s)
      島崎洋次, 真鍋 敬
    • Journal Title

      Biochim. Biophys. Acta 1749

      Pages: 95-101

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Rapid Separation of Microorganism by Quartz Microchip Capillary Electrophoresis2005

    • Author(s)
      Tomoyoshi Shintani, Masaki Torimura, Hiroaki Sato, Hiroaki Tao, Takashi Manabe
    • Journal Title

      Anal.Sci. 21

      Pages: 57-60

    • NAID

      10014095894

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Alkaline Cleavage of Covalent Bonds in Chicken Insulin and Bovine alpha-Lactalbumin Analyzed by Matrix Assisted Laser Desorption/Ionization Mass Spectrometry2005

    • Author(s)
      Takashi Manabe, Ya Jin
    • Journal Title

      Electrophoresis 26

      Pages: 257-267

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] High-efficiency protein extraction from polyacrylamide gels for molecular mass measurement by matrix-assisted laser desorption/ionization-time of flight-mass spectrometry2005

    • Author(s)
      Ya Jin, Takashi Manabe
    • Journal Title

      Electrophoresis 26

      Pages: 1019-1028

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Direct targeting of human plasma for matrix-assisted laser desorption/ionization and analysis of plasma proteins by time of flight-mass spectrometry2005

    • Author(s)
      Ya Jin, Takashi Manabe
    • Journal Title

      Electrophoresis 26

      Pages: 2823-2834

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Detection of activity and mass spectrometric identification of mouse liver caboxylesterase and aldehyde dehydrogenase separated by non-denaturing two-dimensional electrophoresis after extraction with detergents2005

    • Author(s)
      Yoji Shimazaki, Takashi Manabe
    • Journal Title

      Biochim.Biophys.Acta 1749

      Pages: 95-101

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] High-efficiency protein extraction from polyacrylamide gels for molecular mass measurement by matrix-assisted laser desorption/ionization-time of flight mass spectrometry2005

    • Author(s)
      金 亜, 真鍋 敬
    • Journal Title

      Electrophoresis 26・6

      Pages: 1019-1028

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Detection of activity and mass spectrometric identification of mouse liver carboxylesterase and aldehyde dehydrogenase separated by non-denaturing two-dimensional electrophoresis after extraction with detergents2005

    • Author(s)
      島崎 洋次, 真鍋 敬
    • Journal Title

      Biochim.Biophys.Acta 1749・1

      Pages: 95-101

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Direct targeting of human plasma for matrix-assisted laser desorption/ionization and analysis of plasma proteins by time of flight-mass spectrometry2005

    • Author(s)
      金 亜, 真鍋 敬
    • Journal Title

      Electrophoresis 26・14

      Pages: 2823-2834

    • Related Report
      2005 Annual Research Report
  • [Journal Article] 二次元電気泳動法による蛋白質の同定2005

    • Author(s)
      真鍋 敬
    • Journal Title

      検査と技術 33・3

      Pages: 235-240

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Alkaline Cleavage of Covalent Bonds in Chicken Insulin and Bovine alpha-Lactalbumin Analyzed by Matrix Assisted Laser Desorption/Ionization Mass Spectrometry2005

    • Author(s)
      真鍋 敬, 金 亜
    • Journal Title

      Electrophoresis 26・1

      Pages: 257-267

    • Related Report
      2004 Annual Research Report
  • [Journal Article] High-efficiency protein extraction from polyacrylamide gels for molecular mass measurement by matrix-assisted laser desorption/ionization-time of flight mass spectrometry2005

    • Author(s)
      金 亜, 真鍋 敬
    • Journal Title

      Electrophoresis 26・6(印刷中)

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Analysis of Esterase and Transferase Activities in Cytosol Proteins from the Bovine Retina by Uning Microscale Non-Denaturing Two-Dimensional Electrophoresis2004

    • Author(s)
      島崎洋次, 平加容子, 上杉美夏, 真鍋 敬
    • Journal Title

      Biochim. Biophys. Acta 1696

      Pages: 51-57

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Nondenaturing Two-dimensional Electrophoresis Enzyme Profile Involving Activity and Sequence Structure of Cytosol Proteins from Mouse Liver2004

    • Author(s)
      島崎洋次, 菅原 由貴, 真鍋 敬
    • Journal Title

      Proteomics 4

      Pages: 1406-1411

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Concentration and Separation of Low-Abundant Proteins in Human Plasma by Ammonium Sulfate Fractionation Followed by a Three-Step Electrophoresis Technique2004

    • Author(s)
      向井 準, 真鍋 敬
    • Journal Title

      J. Electrophoresis 48

      Pages: 59-66

    • NAID

      130003607324

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Proteome Analysis of Cashmere2004

    • Author(s)
      横濱道成, 増田 豪, 天野 卓, 平山弘樹, 真鍋 敬
    • Journal Title

      Anim. Sci. J. 75

      Pages: 401-405

    • NAID

      10018625347

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Simultaneous Analysis of Esterase and Transferase Activities in Cytosol Proteins from the Bovine Retina by Uning Microscale Non-Denaturing Two-Dimensional Electrophoresis2004

    • Author(s)
      Yoji Shimazaki, Yoko Hiraka, Minatsu Uesugi, Takashi Manabe
    • Journal Title

      Biochim.Biophys.Acta 1696

      Pages: 51-57

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Nondenaturing Two-dimensional Electrophoresis Enzyme Profile Involving Activity and Sequence Structure of Cytosol Proteins from Mouse Liver2004

    • Author(s)
      Yoji Shimazaki, Yuki Sugawara, Takashi Manabe
    • Journal Title

      Proteomics 4

      Pages: 1406-1411

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Concentration and Separation of Low-Abundant Proteins in Human Plasma by Ammonium Sulfate Fractionation Followed by a Three-Step Electrophoresis Technique2004

    • Author(s)
      Jun Mukai, Takashi Manabe
    • Journal Title

      J.Electrophoresis 48

      Pages: 59-66

    • NAID

      130003607324

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Proteome Analysis of Cashmere2004

    • Author(s)
      Michinari Yokohama, Takeshi Masuda, Takashi Amano, Hiroki Hirayama, Takashi Manabe
    • Journal Title

      Anim.Sci.J. 75

      Pages: 401-405

    • NAID

      10018625347

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Nondenaturing Two-dimensional Electrophoresis Enzyme Profile Involving Activity and Sequence Structure of Cytosol Proteins from Mouse Liver2004

    • Author(s)
      島崎 洋次, 菅原 由貴, 真鍋 敬
    • Journal Title

      Proteomics 4・5

      Pages: 406-411

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Concentration and Separation of Low-Abundant Proteins in Human Plasma by Ammonium Sulfate Fractionation Followed by a Three-Step Electrophoresis Technique2004

    • Author(s)
      向井 準, 真鍋 敬
    • Journal Title

      J.Electrophoresis 48・2

      Pages: 59-66

    • NAID

      130003607324

    • Related Report
      2004 Annual Research Report
  • [Book] タンパク質研究なるほどQ&A2005

    • Author(s)
      真鍋 敬ほか36名
    • Publisher
      羊土社
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Annual Research Report 2005 Final Research Report Summary
  • [Book] Protein Analysis Q and A (in Japanese)2005

    • Author(s)
      Takashi Manabe
    • Publisher
      Youdosha
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Book] 先端の分析法2004

    • Author(s)
      真鍋 敬ほか161名
    • Publisher
      エヌ・ティー・エス
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Book] 機器分析化学2004

    • Author(s)
      真鍋 敬ほか61名
    • Publisher
      朝倉書店
    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Book] The Most Advanced Analytical Methods (in Japanese)2004

    • Author(s)
      Takashi Manabe
    • Publisher
      N.T.S.
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Book] Instrumental Analysis (in Japanese)2004

    • Author(s)
      Takashi Manabe
    • Publisher
      Asakura Shoten
    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary

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Published: 2004-04-01   Modified: 2016-04-21  

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