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Analysis of the proteolytic events on cell surface through proteomic approach

Research Project

Project/Area Number 15209011
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section一般
Research Field General medical chemistry
Research InstitutionUniversity of Tokyo

Principal Investigator

SEIKI Motoharu  The University of Tokyo, Institute of Medical Science, Professor, 医科学研究所, 教授 (10154634)

Co-Investigator(Kenkyū-buntansha) YANA Ikuo  The University of Tokyo, Institute of Medical Science, Lecturer, 医科学研究所, 講師 (70332583)
IZUMI Tomonori  The University of Tokyo, Institute of Medical Science, Research Scientist(Donation Laboratory), 医科学研究所, 寄付研究部門教員(常勤形態) (00261694)
Project Period (FY) 2003 – 2005
Project Status Completed (Fiscal Year 2005)
Budget Amount *help
¥49,920,000 (Direct Cost: ¥38,400,000、Indirect Cost: ¥11,520,000)
Fiscal Year 2005: ¥16,250,000 (Direct Cost: ¥12,500,000、Indirect Cost: ¥3,750,000)
Fiscal Year 2004: ¥16,250,000 (Direct Cost: ¥12,500,000、Indirect Cost: ¥3,750,000)
Fiscal Year 2003: ¥17,420,000 (Direct Cost: ¥13,400,000、Indirect Cost: ¥4,020,000)
KeywordsMembrane-type MMP / Integrins / Menbrane proteins / Proteomic analysis / MT1-MMP / 浸潤装置 / がん細胞 / プロテアーゼ / 細胞接着分子 / Matrix metalloproteinase / Proteolysis / Membrane proteins / proteomics / nano-LC-MS / MS
Research Abstract

We established a transfected human squemous carcinoma A431 cell line m which expression of FLAG-tagged MT1-MMP or FIAG-tagged integrins beta-1 is inducible. We set up conditions to isolate the FLAG-tagged MT1-MMP including various associating molecules using different types of detergents. We performed systemic analysis of the proteins in the complex. Five secreted proteins, 64 membrane proteins, 87 cytoplasmic proteins, and 7 unknown proteins were identified Then, we examined 18 membrane proteins whether they can be substrates of MT1-MMP by expressing them in COS-1 cells. Nine of them were found to be cleaved by the expression of MT1-MMP generating processed fragments. One of the processed proteins has been reported to be overexpressed many tumors and be shed by metalloproteinases. We confirmed that this protein also cleaved by endogenous MT1-MMP in human tumor cell lines that express MT1-MMP. Thus, we believe this approach is useful to identify MT1-MMP substrates and regulators by associating with MT1-MMP We are trying to know the functions of the substrates and associating proteins by the knock-down experiments using siRNA. The association of the identified proteins with MT1-MMP is further tested using cross-linkers. We are continuing analysis of integrin-assoaating proteins as well.

Report

(4 results)
  • 2005 Annual Research Report   Final Research Report Summary
  • 2004 Annual Research Report
  • 2003 Annual Research Report
  • Research Products

    (28 results)

All 2005 2004 Other

All Journal Article (22 results) Publications (6 results)

  • [Journal Article] Palmitoyalion at Cys574 is essential for MT1-MMP to promote cell migration2005

    • Author(s)
      Anilkumar, N, Seiki M, 他4名
    • Journal Title

      FASEB J 19・10

      Pages: 1326-8

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Impaired alveolization in mice deficient in membrane-type matrix metalloproteinase 1 (MT1-MMP)2005

    • Author(s)
      Irie, K, Seiki, M, 他5名
    • Journal Title

      Med Mol Morphol 38・1

      Pages: 43-6

    • NAID

      10018374513

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Matrix metalloproteinase expression and function during fin regeneration in zebrafish : analysis of MT1-MMP, MMP2 and TIMP22005

    • Author(s)
      Bai, S, Seiki, M, 他8名
    • Journal Title

      Matrix Biol 24・4

      Pages: 247-60

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Competitive disruption of the tumor-promoting function of membrane type 1 matrix metalloproteinase/matrix metalloproteinase-14 in vivo2005

    • Author(s)
      NoNaka, T, Yana, I, Seiki, M
    • Journal Title

      Mol Cancer Ther 4・8

      Pages: 1157-66

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] CD44 binding through the hemopexin-like domain is criticat for its shedding by membrane-type 1 matrix metalloproteinase2005

    • Author(s)
      Suenaga, N, Seiki, M, 他2名
    • Journal Title

      Oncogene 24・5

      Pages: 859-68

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Membrane-type matrix metalloproteinase-1 (MT1-MMP) is a processing enzyme for human laminin gamma 2 chain2005

    • Author(s)
      Koshikawa, N, Seiki, M, 他3名
    • Journal Title

      J Biol Chem. 280・1

      Pages: 88-93

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Palmitoylation at Cys574 is essential for MT1-MMP to promote cell migration2005

    • Author(s)
      Anikumar, N, Seiki, M(four others).
    • Journal Title

      FASEB J Vol.19, No.19

      Pages: 1326-1328

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Impaired alveolization in mice deficient in membrane-type matrix metalloproteinase 1 (MT1-MMP)2005

    • Author(s)
      Irie, K., Seiki, M.(five others).
    • Journal Title

      Med.Mol.Morphol. Vol.38, No.1

      Pages: 43-46

    • NAID

      10018374513

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Matrix metalloproteinase expression and function during fin regeneration in zebrafish : analysis of MT1-MMP, MMP2 and TIMP22005

    • Author(s)
      Bai, S., Seiki, M.(eight others).
    • Journal Title

      Matrix Biol. Vol.24, No.4

      Pages: 247-260

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Competitive disruption of the tumor-promoting function of membrane type 1 matrix metalloproteinase/matrix metalloproteinase-14 in vivo2005

    • Author(s)
      Nonaka, T., Seiki, M.(three others).
    • Journal Title

      Mol.Can.Ther. Vol.4, No.8

      Pages: 1157-1166

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] CD44 binding through the hemopexin-like domain is critical for its shedding by membrane-type 1 matrix metalloproteinase2005

    • Author(s)
      Suenaga, N., Seiki, M.(two others).
    • Journal Title

      Oncogene Vol.24, No.5

      Pages: 859-868

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Membrane-type matrix metalloproteinase-1 (MT1-MMP) is a processing enzyme for human laminin gamma 2 chain2005

    • Author(s)
      Koshikawa, N., Seiki, M.(three others).
    • Journal Title

      J.Biol.Chem. Vol.280, No.1

      Pages: 88-93

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2005 Final Research Report Summary
  • [Journal Article] Palmitoylation at Cys574 is essential for MT1-MMP to promote cell migration2005

    • Author(s)
      Anilkumar, N. 他5名
    • Journal Title

      FASEB J 19・10

      Pages: 1326-1328

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Impaired alveolization in mice deficient in membrane-type matrix metalloproteinase 1 (MT1-MMP)2005

    • Author(s)
      Irie, K. 他6名
    • Journal Title

      Med Mol Morphol 38・1

      Pages: 43-46

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Matrix metalloproteinase expression and function during fin regeneration in zebrafish : analysis of MT1-MMP, MMP2 and TIMP22005

    • Author(s)
      Bai, S. 他9名
    • Journal Title

      Matrix Biol 24・4

      Pages: 247-260

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Competitive disruption of the tumor-promoting function of membrane type 1 matrix metalloproteinase/matrix metalloproteinase-14 in vivo2005

    • Author(s)
      Nonaka, T. 他4名
    • Journal Title

      Mol Cancer Ther 4・8

      Pages: 1157-1166

    • Related Report
      2005 Annual Research Report
  • [Journal Article] CD44 binding through the hemopexin-like domain is critical for its shedding by membrane-type 1 matrix metalloproteinase2005

    • Author(s)
      Suenaga, N. 他3名
    • Journal Title

      Oncogene 24・5

      Pages: 859-868

    • Related Report
      2005 Annual Research Report
  • [Journal Article] Membrane-type matrix metalloproteinase-1 (MT1-MMP) is a processing enzyme for human laminin gamma 2 chain2005

    • Author(s)
      Koshikawa, N. 他4名
    • Journal Title

      J Biol Chem. 280・1

      Pages: 88-93

    • Related Report
      2005 Annual Research Report
  • [Journal Article] CD44 binding through the hemopexin-like domain is critical for its shedding by membrane-type 1 matrix metalloroteinase2005

    • Author(s)
      Suenaga, N.他3人
    • Journal Title

      Oncogene 24

      Pages: 859-868

    • Related Report
      2004 Annual Research Report
  • [Journal Article] MTCBP-1 is a new member of the Cupin superfamily. A possible multifunctional protein acting as an invasion suppressor down-regulated in tumors2004

    • Author(s)
      Uekita, T.他7人
    • Journal Title

      J Biol Chem 279

      Pages: 12734-12743

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Absence of mechanical allodynia and Abeta-fiber sprouting after sciatic nerve injury in mice lacking membrane-type 5 matrix metalloproteinase2004

    • Author(s)
      Komori, K.他7人
    • Journal Title

      FEBS Lett. 557

      Pages: 125-128

    • Related Report
      2004 Annual Research Report
  • [Journal Article] Constitutive and induced CD44 shedding by ADAM-like proteases and membrane-type 1 matrix metalloproteinase2004

    • Author(s)
      Nakamura, H.他7人
    • Journal Title

      Cancer Res. 64

      Pages: 876-882

    • Related Report
      2004 Annual Research Report
  • [Publications] Nakamura H., et al.: "Constitutive and induced CD44 shedding by ADAM-like proteases and membrane-type matrix."Cancer Res.. 64. 876-882 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Uekita T., et al.: "Membrane-type 1 matrix metalloproteinase cytoplasmic tail binding protein-1 (MTCBP-1) is a new member of the Cupin superfamily : A possible multifunctional protein acting as an invasion."J.Biol.Chem.. (in press). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Komori K., et al.: "Absence of mechanical allodynia and Abeta-fiber sprouting after sciatic nerve injury in mice lacking membrane-type 5 matrix metalloproteinase."FEBS Lett.. 557. 125-128 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Matsuda A., et al.: "Clusterin, an abundant serum factor, is a possible negative regulator of MT6-MMP/MMP-25 produced by neutrophils."J.Biol.Chem.. 278. 36350-36357 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Matsuki H., et al.: "Monoclonal antibodies with defined recognition sequences in the stem region of CD44 : detection of differential glycosylation of CD44 between tumor and stromal cells in tissue."Cancer Res.. 63. 8278-8283 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] Ueda J., et al.: "Sequence-specific silencing of MT1-MMP expression suppresses tumor cell migration and invasion : importance of MT1-MMP as a therapeutic target for invasive tumors."Oncogene. 22. 8716-8722 (2003)

    • Related Report
      2003 Annual Research Report

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Published: 2003-04-01   Modified: 2016-04-21  

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