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1991 Fiscal Year Final Research Report Summary

Mechanism of Activation of Signaling Enzymes in Inositol Lipid Metabolism Following Ligation of Cell Surface Receptors in T Lymphocytes

Research Project

Project/Area Number 02807029
Research Category

Grant-in-Aid for General Scientific Research (C)

Allocation TypeSingle-year Grants
Research Field General medical chemistry
Research InstitutionSapporo Medical College

Principal Investigator

SASAKI Terukatsu  Sapporo Medical College, Cancer Research Institute, Professor, 癌研究所, 教授 (00045494)

Co-Investigator(Kenkyū-buntansha) YAMASHITA Yumi  Sapporo Medical College, Cancer Research Institute, Res. Associate, 癌研究所, 研究技師
ISHINO Masaho  Sapporo Medical College, Cancer Research Institute, Instructor, 癌研究所, 助手 (30232325)
SASAKI Hiroko  Sapporo Medical College, Cancer Research Institute, Assistant Prof., 癌研究所, 講師 (60045424)
Project Period (FY) 1990 – 1991
KeywordsT cell receptor / protein tyrosine kinase / tyrosine phosphorylation / src homology region 2 / phospholipase C / recombinant baculovirus / cytoplasmic free Ca^<2+> concentration
Research Abstract

1. An increase in the cytoplasmic free Ca^<2+> concentration([Ca^<2+>]i)occurs on ligation with antibody of the T cell receptor (TCR) / CD3 complex of a human T cell leukemia line, Jurkat. It was found that the[Ca^<2+>]i response is inhibited by staurosporine(Stsp). Stsp markedly inhibited tyrosine phosphorylation of proteins found on ligation of the TCR/CD3 complex. Stsp had no effect on the[Ca^<2+>]i response in fibroblasts following the stimulation of cell surface receptors coupled to G-proteins, while the[Ca^<2+>]i response to the activation of receptor tyrosine kinase was inhibited by Stsp. These results indicate that a protein tyrosine kinase is involved in the[Ca^<2+>]i response to ligation of the TCR/CD3 complex.
2. The ligation of the TCR/CD3 complex induces tyrosine phosphorylation of the zeta chain of the complex. Tyrosine-phosphorylated zeta chain may function as a binding site of proteins with src homology region 2(SH2)involved in the signal transduction in T lymphocytes. In order to screen such proteins, rat zeta chain and human p59^<fyn> are co-expressed in baculovirus system. The expression of zeta chain was confirmed by immunoblotting with anti zeta peptide(residues 110-122)antibody. Simultaneous expression of zeta chain and p59^<fyn> in SF9 cells resulted in the tyrosinephosphorylation of zeta chain. Screening for the proteins containing SH2 region are in progress by the use of lambdagt11 library of cDNA from Jurkat cells and the phosphorylated zeta chain, labeled with[ ^<32>P]phosphate at tyrosine residues, as a probe.
3. Binding specificity of SH2 regions present in phospholipase Cgamma(PLCgamma)and nonreceptor protein tyrosine kinases, p56^<1ck> and p59^<fyn>, is studied. The SH2 regions of these proteins were expressed in E. coli as fusion proteins with the trpE gene product and affinity column of the SH2 regions were prepared. The SH2 region of PLC had different binding specificity from those of the SH2 regions of p56^<1ck> and p59^<fyn>.

  • Research Products

    (11 results)

All Other

All Publications (11 results)

  • [Publications] Y.Yamashita: "Suppression by Staurosporine of Ca^<2+>-mobilization triggered by ligation of antigen-specific receptors on T and B lymphocytes." FEBS Lett.288. 46-50 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Takahashi: "Preparation of anti-T idiotype monoclonal antibody reacting with human T leukaemic cell lines and with small percentage of peripheral T lymphocytes." Clin.exp.Imuunol.82. 590-595 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] H.Hasegawa-Sasaki: "Mechanism of phospholipase C activation through the T cell receptor/ CD3 complex." Tumor Res.25. 37-54 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] T.Endo: "Difference in the Ca^<2+> responses of various human T cell lines to stimulation of the T cell receptor/CD3 complex with monoclonal antibodies." Tumor Res.25. 55-62 (1990)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 佐々木 輝捷: "プロテインチロシンキナ-ゼによるイノシト-ルリン脂質代謝酸素の活性化" 代謝(Vol.28臨時増刊号). 28. 69-73 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 佐々木 輝捷: "Tリンパ球の活性化とイノシト-ルリン脂質代謝" 蛋白質核酸酵素. 36. 235-239 (1991)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 佐々木 輝捷(分担執筆): "「ニュ-ロサイエンス講座」第5巻4.9受容体プロテインチロシンキナ-ゼ系によるホスホリパ-ゼCの活性化" 廣川書店,

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Y. Yamashita et al.: "Suppression by staurosporine of Ca^<2+>-mobilization triggered by ligation of antigen-specific receptors on T and B lymphocytes." FEBS Lett.288. 46-50 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T. Takahashi et al.: "Preparation of anti-T idiotype monoclonal antibody reacting with human T leukaemic cell lines and with small percentage of peripheral T lymphocytes." Clin. exp. Immunol.82. 590-595 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] H. Hasegawa-Sasaki et al.: "Mechanism of phospholipase C activation through the T cell receptor/ CD3 complex." Tumor Res.25. 37-54 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] T. Endo et al.: "Difference in the Ca^<2+> responses of various human T cell lines to stimulation of the T cell receptor/CD3 complex with monoclonal antibodies." Tumor Res.25. 55-62 (1990)

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

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Published: 1993-03-16  

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