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The function of DG kinase, a lipid phosphorylating enzyme having both EF-hands and zinc fingers.

Research Project

Project/Area Number 02454157
Research Category

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

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

Principal Investigator

KANOH Hideo  Sapporo Medical College, Faculty of Medicine, Professor, 医学部, 教授 (70045475)

Co-Investigator(Kenkyū-buntansha) IMAI Shin-ichi  Sapporo Medical College, Faculty of medicine, Research Associate, 医学部, 助手 (20213209)
SAKANE Fumio  Sapporo Medical College, Faculty of medicine, Research Associate, 医学部, 助手 (10183815)
YAMADA Keiko  Sapporo Medical College, Faculty of medicine, Assistant Professor, 医学部, 講師 (80045541)
Project Period (FY) 1990 – 1991
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1991: ¥800,000 (Direct Cost: ¥800,000)
KeywordsDiacylglycerol kinase / EF-hand motif / zinc finger / Calcium-binding protein / cDNA cloning / Signal transduction / ホスファチジン酸 / ジアシルグリセロ-ル / リン脂質合成 / リンパ球
Research Abstract

We have cloned cDNA encoding porcine 80K diacylglycerol kinase (DGK), thus demonstrating the presence of a novel enzyme having both EF-hands and zinc fingers (Sakane, Nature, 1990). Based on this finding we studied the structure-function relationship of this enzyme using mainly molecular genetics.
We showed that the purified enzyme is indeed a high affinity Ca^<2+>-binding protein. In the absence of Ca^<2+> the EF-hand region serves as an autoinhibitory domain by masking a putative phosphatidylserine binding site (s) of this enzyme. Upon Ca^<2+> binding the EF-hands undergoes conformational change that results in the exposure of phospholipid binding site(s). (Sakane, JBC, 1991, Sakane, BBRC, 1991).
We cloned cDNA of the rat homolog of the 80K DGK. This enabled us to study the mode of enzyme expression in detail by in situ hybridization and immunostaining. The 80K DGK and its gene were highly expressed in T-lymphocytes, while in the central nervous system their expression is confined to oligodendrocytes. The mode of mRNA formation was linked to the maturation of myelin tissues. We are currently cloning cDNA coding for neuron-specific DGK using the 80K DGK cDNA as a probe. (Goto, submitted). In this respect, three DGK isozymes were purified from human platelets, and they were shown to be distinct from the 80K isozyme (Yada, JBC, 1990).
At present the transcriptional control mechanism of the 80K enzyme is being studied using human gene already cloned. The function of zinc finger is also studied by expressing the mutated enzyme in Baculovirus/Sf9 cell system. The function of the 80K enzyme in T-lymphocytes is investigated by immunocytochemical methods.

Report

(3 results)
  • 1991 Annual Research Report   Final Research Report Summary
  • 1990 Annual Research Report
  • Research Products

    (54 results)

All Other

All Publications (54 results)

  • [Publications] Kanoh,Hideo: "Diacylglycerol Kinase:a key modulator of signal transduction?" Trends in Biochemical Sciences. 15. 47-50 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Sakane,Fumio: "Porcine diacylglycerol kinase sequence has zinc finger and E-F hand motifs." Nature. 344. 345-348 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Yada,Yukihiro: "Purification and characterization of cytosolic diacylglycerol kinases of human platelets." Journal of Biological Chemistry. 265. 19237-19243 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Imaizumi,Toshio: "Non-heparin-binding endothelial cell growth factor from bovine omentum." Experimental Cell Research. 187. 292-298 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Sakane,Fumio: "porcine 80 kDa diacylglycerol kinase is a calcium-binding and calciumuphospholipid dependent enzyme and undergoes calcium dependent translocation." Journal of Biological Chemistry. 266. 7096-7100 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Sakane,Fumio: "The regulatory role of EF-hand motifs of pig 80K diacylglycerol Kinase as assessed using truncation and deletion mutaznts." Biochemical and Biophysical Research Communications. 181. 1015-1021 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Goto,Kaoru: "Specific expression of 80 kDa-diacylglycerol kinase and its gene in oligodendrocytes of the rat brain." Journal of Neuroscience.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 加納 英雄: "カルモデュリン様構造と亜鉛フィンガ-をもつジグリセリドキナ-ゼ" 蛋白質核酸酵素. 35. 1558-1563 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 加納 英雄: "ジアシルグリセロ-ルキナ-ゼ" 実験医学. 8. 2043-2048 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 坂根 郁夫: "ジアシルグリセロ-ルキナ-ゼの一次構造と活性調節" 蛋白質核酸酵素. 36. 290-298 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 加納 英雄: "ジグリセリドキナ-ゼーカルシウムセンサ-をもつ情報伝達制御因子" 代謝. 28. 101-106 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 加納 英雄: "リン脂質合成酵素研究の新しい展開" New Lipids.(日本油脂株式会社). 3. 1-6 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Kanoh,Hideo(ed.by E.A.Dennis & D.E.vance): "Diacylglycerol kinase isozymes from brain and lymphoid tissues.(Methods in Enzymology,Vol.209)" Academic Press,San Diego,U.S.A, 162-172(11) (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 山田 恵子: "ジアシルグリセロ-ルキナ-ゼ(新生化学実験講座.第4巻)" 東京化学同人, 355-366(12) (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 加納 英雄: "DGキナ-ゼNewroscience講座V,セカンドメッセンジャ-と産生酵素(竹縄,野村編)" 広川書店,

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 坂根 郁夫: "DGキナ-ゼ 生物薬科学講座IV 生体膜の構造と機能(石橋,寺田編)" 広川書店,

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Kanoh, H.,: "Diacylglycerol kinase : a key modulator of signal transduction?" Trends Biochem. Sci.15. 47-50 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Sakane, F.: "Porcine diacylglycerol kinase sequence has zinc finger and E-F hand motifs." Nature. 344. 345-348 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Yada, Y.: "Purification and characterization of cytosolic diacylglycerol kinases of human platelets." J. Biol. Chem.265. 19237-19243 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Imaizumi, T.: "Non-heparin-binding endothelial cell growth factor from bovine omentum." Exp. Cell Res.187. 292-298 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Sakane, F.: "Porcine 80-kDa diacylglycerol kinase is a calcium-binding and calcium/phospholipid dependent enzyme and undergoes calcium dependent translocation." J. Biol. Chem.266. 7096-7100 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Sakane, F.: "The regulatory role of EF-hand motifs of pig 80K diacylglycerol kinase as assessed using truncation and deletion mutant." Biochem. Biophys. Res. Commun.181. 1015-1021 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Kanoh, H.: "Diacylglycerol kinase isozymes from brain and lymphoid tissues." Methods Enzymol.209. 162-172 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Goto, K.: "Specific expression of 80-kDa diacylglycerol kinase and its gene in oligodendrocytes of the rat brain."

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Kanoh,Hideo: "Diacylglycerol kinase:a key modulator of signal transduction?" Trends in Biochemical Sciences. 15. 47-50 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] Sakane,Fumio: "Porcine diacylglycerol kinase sequence has zinc finger and EーF hand motifs" Nature. 344. 345-348 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] Yada,Yukihiro: "Purification and characterization of cytosolic diacylglycerol kinases of human platelets" Journal of Biological Chemistry. 265. 19237-19243 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] Imaizumi,Toshio: "Nonーheparinーbinding endothelial cell growth factor from bovine omentum" Experimental Cell Research. 187. 292-298 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] Sakane,Fumio: "Porcine 80 kDa diacylglycerol kinase is a calciumーbindind and calcium/phospholipid dependent enzyme and undergoes calcium dependent translocation" Journal of Biological Chemistry. 266. 7096-7100 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Sakane,Fumio: "The regulatory role of EFーhand motifs of pig 80K diacylglycerol kinase as assessed using truncation and deletion mutants" Biochemical and Biophysical Research Communications. 181. 1015-1021 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Goto,Kaoru: "Specific expression of 80 kDaーdiacylglycerol kinase and its gene in oligodendrocytes of the rat brain." Journal of Neuroscience.

    • Related Report
      1991 Annual Research Report
  • [Publications] 加納 英雄: "カルモデュリン様構造と亜鉛フィンガ-をもつジグリセリドキナ-ゼ" 蛋白質核酸酵素. 35. 1558-1563 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] 加納 英雄: "ジアシルグリセロ-ルキナ-ゼ" 実験医学. 8. 2043-2048 (1990)

    • Related Report
      1991 Annual Research Report
  • [Publications] 坂根 郁夫: "ジアシルグリセロ-ルキナ-ゼの一次構造と活性調節" 蛋白質核酸酵素. 36. 290-298 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 加納 英雄: "ジグリセリドキナ-ゼーカルシウムセンサ-をもつ情報伝達制御因子" 代謝. 28. 101-106 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 加納 英雄: "リン脂質合成酵素研究の新しい展開" New Lipids.(日本油脂株式会社). 3. 1-6 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] Kanoh,Hideo(ed.by E.A.Dennis & D.E.Vance): "Diacylglycerol kinase isozymes from brain and lymphoid tissues.(Methods in Enzymology,Vol.209)" Academic Press,San Diego,U.S.A., 162-172(11) (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] 山田 恵子: "ジアシルグリセロ-ルキナ-ゼ(新生化学実験講座.第4巻)" 東京化学同人, 355-366(12) (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 加納 英雄: "DGキナ-ゼ Neurosuence講座V,セカンドメッセンジャ-と産生酵素(竹縄,野村編)" 広川書店,

    • Related Report
      1991 Annual Research Report
  • [Publications] 坂根 郁夫: "DGキナ-ゼ 生物薬科学講座IV 生体膜の構造と機能(石橋,寺田編)" 広川書店,

    • Related Report
      1991 Annual Research Report
  • [Publications] 加納 英雄: "Diacylglycerol kinase:a key modulator of signal transduction?" Trends in Biochemical Sciences. 15. 47-50 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 坂根 郁夫: "Porcine diacylglycerol kinase sequence has zinc finger and EーF hand motifs" Nature. 344. 345-348 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 矢田 幸博: "Purification and characterization of cytosolic diacylglycerol Kinases of human platelets" Journal of Biological Chemistry. 265. 19237-19243 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 今泉 俊雄: "Nonーheparinーbinding endothelial cell growth factor from bovine omentum" Experimental Cell Research. 187. 292-298 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 坂根 郁夫: "Porcine 80 kDa diacylglycerol kinase is a calcium binding and calcium/phospholipid dependent enzyme and undergoes calcium dependdent translocation." Journal of Biological Chemistry. (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] 山田 恵子: "Effect of sphingosine on diacylglycerol kinase phosphorylating endogenous and exogenous diacylglycerols in human T lymphocytes" J.Biol.Chem.

    • Related Report
      1990 Annual Research Report
  • [Publications] 加納 英雄: "カルモデュリン様構造と亜鉛フィンガ-をもつジグリセリドキナ-ゼ" 蛋白質核酸酵素. 35. 1558-1563 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 加納 英雄: "DGキナ-ゼ" 実験医学. 8. 2043-2048 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 坂根 郁夫: "ジアルシルグリセロ-ルキナ-ゼの1次構造と機能調節" 蛋白質核酸酵素. 36. 290-298 (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] 加納 英雄: "ジグリセリドキナ-ゼーカルシウムセンサ-をもつ情報伝達制御因子" 代謝. (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] 加納 英雄(編集者E.A.Dennis & D.E.Vance): "Diacylglycerol kinase isozymes from brain and lymphoid tissues.(Methods in Enzymology)" Academic Press,Florida,U.S.A., (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] 山田 恵子: "ジアシルグリセロ-ルキナ-ゼ(新生化学実験講座)" 東京化学同人社, (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] 加納 英雄: "DGキナ-ゼ(ニュ-ロサイエンス講座)" 廣川書店, (1991)

    • Related Report
      1990 Annual Research Report
  • [Publications] 坂根 郁夫: "DGキナ-ゼ(生物薬科学実験講座)" 廣川書店, (1991)

    • Related Report
      1990 Annual Research Report

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Published: 1991-04-01   Modified: 2016-04-21  

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