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2004 Fiscal Year Final Research Report Summary

Discovery of protein-to-protein interactions in platelets by a novel methods.

Research Project

Project/Area Number 15590991
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Hematology
Research InstitutionHOKKAIDO UNIVERSITY

Principal Investigator

ODA Atsushi  Hokkaido Univ., Grad.School of Med., Lecturer, 大学院・医学研究科, 助教授 (50255436)

Co-Investigator(Kenkyū-buntansha) FUJITA Hiroyoshi  Hokkaido Univ., Grad.School of Med., Professor, 大学院・医学研究科, 教授 (60142931)
Project Period (FY) 2003 – 2004
Keywordsplatelet / signal transduction / mass spectrometry / actin / rac
Research Abstract

Searching for proteins in platelets that can interact with the N-terminal SH3 domain of CrkL (using a combination of a pull-down assay followed by mass spectrometry), we have found that human platelets express DOCK5, a putative Rac guanine nucleotide exchanger, as a CrkL-binding protein. We cloned DOCK5 and raised specific antibodies against the molecule. We overexpressed CrkL, DOCK5, or both in combination in COS7 cells. Overexpressed DOCK5 showed diffuse cytoplasmic distribution. However, when co-expressed with wild-type CrkL, both DOCK5 accumulated at CrkL-induced focal adhesions, suggesting a functional implication of the interaction.
Using specific antibodies against isoforms of WAVE (WASP [Wiskott-Aldrich syndrome protein] family Verprolin-homologous protein, also called Scar), we demonstrated that human platelets express all 3 isoforms. With the use of an in vitro pull-down technique, the src homology 3 (SH3) domain of insulin receptor substrate p53 (IRSp53) precipitated WAVE2 from platelet lysates more efficiently than did profilin I. The opposite was true for WAVE1, and neither precipitated WAVE3, suggesting that WAVE isoforms have different affinities to these ligands, while the SH3 domain of abl binds to all 3 isoforms. By mass spectrometry, we found that the proteins, which reportedly interact with WAVE/Scars, are present in platelets.
Thus, we have effectively employed mass spectrometry to demonstrate hitherto unreported proteins in platelets and their interactions.

  • Research Products

    (12 results)

All 2005 2003

All Journal Article (12 results)

  • [Journal Article] WAVE/Scars in Platelets.2005

    • Author(s)
      Oda A, Fujita H, et al.
    • Journal Title

      Blood 105

      Pages: 3141-3148

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Signal transduction and cellular functions of the TEL/ARG2005

    • Author(s)
      Oda A, Fujita H, et al.
    • Journal Title

      Leukemia 19

      Pages: 603-610

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] WAVE/Scars in platelets2005

    • Author(s)
      Oda A, Miki H, Wada I, Yamaguchi H, Yamazaki D, Suetsugu S, Nakajima M, Nakayama A, Okawa K, Miyazaki H, Matsuno K, Ochs HD, Machesky LM, Fujita H, Takenawa T.
    • Journal Title

      Blood 105

      Pages: 3141-3148

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Signal transduction and cellular functions of the TEL/ARG oncoprotein2005

    • Author(s)
      Okuda K, Oda A, Sato Y, Nakayama A, Fujita H, Sonoda Y, Griffin JD.
    • Journal Title

      Leukemia 19

      Pages: 603-610

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] STI571-resistant KT-1 cells are sensitive to interferon-alpha accompanied by the loss of T-cell protein tyrosine phosphatase and prolonged phosphorylation of Stat1.2003

    • Author(s)
      Shimizu T, Oda A, et al.
    • Journal Title

      Exp Hematol. 31

      Pages: 601-608

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Thrombopoietin upregulates nucleolin mRNA and protein in thrombopoietin-dependent megakaryocytic cell line, UT-7/TPO.2003

    • Author(s)
      Ito T, Oda A, et al.
    • Journal Title

      Mol Cell Biochem. 247

      Pages: 75-82

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Bernard-soulier syndrome with a homozygous 13 base pair deletion in the signal peptide-coding region of the platelet glycoprotein Ibβ gene.2003

    • Author(s)
      Watanabe R, Oda A, et al.
    • Journal Title

      Blood Coagul Fibrinolysis. 14

      Pages: 387-394

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] Functional phenotype of phosphoinositide 3-kinase p85α-null platelets characterized by an impaired response to GP VI stimulation.2003

    • Author(s)
      Watanabe N, Oda A, et al.
    • Journal Title

      Blood 102

      Pages: 541-548

    • Description
      「研究成果報告書概要(和文)」より
  • [Journal Article] STI571-resistant KT-1 cells are sensitive to IFN-alpha accompanied by the loss of TC-PTP phosphatase and prolonged phosphorylation of Stat.2003

    • Author(s)
      Shimizu T, Miyakawa Y, Oda A, Kizaki M, Ikeda Y.
    • Journal Title

      Exp Hematol 31

      Pages: 601-608

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Thrombopoietin upregulates nucleolin mRNA and protein in thrombopoietin-dependent megakaryocytic cell line, UT-7/TPO.2003

    • Author(s)
      Ito T, Fujihara M, Oda A, Wakamoto S, Yamaguchi M, Komatsu N, Miyazaki H, Azuma H, Ikeda H, Ikebuchi K.
    • Journal Title

      Mol Cell Blochem. 247

      Pages: 75-82

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Bernard-Soulier syndrome with a homozygous 13-bp deletion in the signal peptide-coding region of platelet glycoprotein Ibβ gene.2003

    • Author(s)
      Watanabe R, Ishabshi T, Saitoh Y, Shichishima T, Maruyama Y, Enomoto Y, Handa M, Oda A, Ambo H, Murata M, Ikeda Y.
    • Journal Title

      B od Coagulation & Fibrinolysis 14

      Pages: 387-394

    • Description
      「研究成果報告書概要(欧文)」より
  • [Journal Article] Functional phenotype of phosphoinositide 3-kinase p85alpha null platelets characterized by an impaired response to GP VI stimulation.2003

    • Author(s)
      Watanabe N, Hideaki Nakamrna H, Suzuki H, Oda A, Matsubara M, Moroi M, Terauchi Y, Takashi Kadowaki T, Suzuki, Shigeo Koyasu S, Ikeda Y, Handa M.
    • Journal Title

      Blood 102

      Pages: 541-548

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2006-07-11  

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