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Study of Mucolipidosis by analysis of phosphorylation enzyme on carbohydrate moiety of lysosomal enzyme

Research Project

Project/Area Number 02670420
Research Category

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

Allocation TypeSingle-year Grants
Research Field Pediatrics
Research InstitutionHokkaido University

Principal Investigator

GASA Shinsei  Hokkaido University School of Medicine, Associated Professor, 医学部, 助教授 (10142712)

Co-Investigator(Kenkyū-buntansha) FUJITA Michiyasu  Hokkaido University School of Dental, Lecturer, 歯学部, 講師 (10181372)
Project Period (FY) 1990 – 1991
Project Status Completed (Fiscal Year 1991)
Budget Amount *help
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1991: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1990: ¥1,600,000 (Direct Cost: ¥1,600,000)
KeywordsSugar Phosphorvlation / Lysosomal enzyme / Mucolipidosis / I-cell disease / Iーcell液 / 糖鎖リン酸化 / Nーアセチルグルコサミンー1ーリン酸転移酵素
Research Abstract

UDP-GlcNAc : lysosmal enzyme GlcNAc 1-phosphotransferase, which defects or partially expresses the activity in Mucolipidosis type II (I-cell disease) or type III, respectively, has not yet been analyzed for the biochemical property, since the enzyme is much less expressed and unstable in many mammalian tissues such as liver, kidney, etc. Furthermore the enzyme requires[beta -^<32>P]UDP-GlcNAc for assay of the activity as a donor, which is not available commercially and needs to be synthesized. These disadvantages, therefore, delay to investigate above diseases in detail. This project aims to analyze immunologically the enzyme protein and further clarify a mechanism of the defect in the Mucolipidosis after purification of the human enzyme.
(1991) 1. The activity of the enzyme was much higher in human leukemic' enzyme and the serum, and human hepatocellular carcinoma tissue as compared to their normal control. Furthermore, rat fibrosarcoma cell KMT-17, established in this university, and the ascites cells demonstrated the highest activity, suggesting better source of the enzyme to purify. 2. The enzyme has been purified from rat liver Golgi membrane by us, and poly clonal antibody against the enzyme has been prepared by rabbit. However, the antibody has not reacted immunologically to human enzyme, showing different protein between rat and human (in preparation to be submitted).
(1992) 1. The purification methods of the rat enzyme has been applied for the human enzyme, however, resulting low purification fold (10^4-fold), since fresh or unfrozen tissue from human has not been available. 2. Since the enzyme catalyzes the phosphorylation to mannan from UDP-GlcNAc, the analogues of these substrates might be employed as a photoaffinity probe. In fact, azidosalycilate derivatives of[beta -^<32>P]UDP-GlcNH_2 and mannan has been synthesized)and the derivatives are appling for detection of the enzyme in several cells.

Report

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

    (27 results)

All Other

All Publications (27 results)

  • [Publications] Y.Uehara: "Increased N-acetylglucosamine-l-phosphotransferase activty in sera from patients with leukemia" Jpn.J.Cancer Res.82. 82-85 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] M.Ohhira: "Elevated carbohydrate phosphotransferase activity in human hepatoma and phosphorylation of cathepsin D" Br.J.Cancer. 63. 905-908 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] S.Gasa: "Elevated serum level of glycolipid sulfotransferase in patients with hepatocellular carcinoma" Cancer Lett.59. 19-24 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] N.Sakakibara: "Distinctive glycolipid patterns in Wilms' tumor and renal cell carcinoma" Cancer Lett.57. 187-192 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] S.Gasa: "Intracellular targentting signal and disease-I-cell disease" BIOmedica. 7. 135-138 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] J.Tanaka: "Characterization of the subunits and sugar moiety of human placental and leukemic -glucuronidase" Biol.Chem.Hoppe-Seyler. 373. 57-62 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 賀佐 伸省: "新生化学実験講座" 東京化学同人, 4 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] 賀佐 伸省: "タンパク質化学" 5 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Y. Uehara: "Increased N-acetylglucosamine-1-phosphotransferase activity in sera from patients with leukemia" Jpn. J. Cancer Res.82. 82-85 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] M. Ohhira: "Elevated carbohydrate phosphotransferase activity in human hepatoma and phosphorylation of cathepsin D" Br. J. Cancer. 63. 905-908 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Gasa: "Elevated serum level of glycolipid sulfotransferase in patients with hepatocellular carcinoma" Cancer Lett.59. 19-24 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] N. Sakakibara: "Distinctive glycolipid patterns in Wilms' tumor and renal cell carcinoma" Cancer Lett.57. 187-192 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] S. Gasa: "Intracellular targeting signal and disease---I cell disease" BIOmedica. 7. 135-138 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] J. Tanaka: "Characterization of the subunits and sugar moiety of human placental and leukemic-glucuronidase" Biol. Chem. Hoppe-Seyler. 373. 57-62 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] S. Gasa: Shinseikagakujikken-Kouza. Lysosomal Disease-I-Cell disease, 15 (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] S. Gasa: Seibutsuyakukagaku-jikkenkouza. Sulfatase-Degradation, Lipid, Glycolipid, (1992)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1991 Final Research Report Summary
  • [Publications] Y.Uehara: "Increased Nーacetylglucosamineー1ーPhosphotransferase activity in sera from patients with leukemia" Jpn,J,Cancer Res,. 82. 82-85 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] M.Ohhira: "Elevated carbohydrate phosphotransferase activity in human hepatome and phosphorylation of cathepsin D" Br.J.Cancer. 63. 905-908 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] S.Gasa: "Elevated serum level of glycolipid sulfotransferase in patients with hepatocellular carcinoma" Cancer Lett.59. 19-24 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] 賀佐 伸省: "細胞内局在シグナルと疾患「Iーcell病」" BIOmedica. 7. 135-138 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] N.Sakakibara: "Distinctive glycolipid patterns in Wilms′tumor and renal cell carcinoma" Cancer Lett.57. 187-192 (1991)

    • Related Report
      1991 Annual Research Report
  • [Publications] J.Tanaka: "Characterization of the subunits and sugar moiety of human placental and leukemic βーglucuronidase" Biol.Chem.HoppeーSeyler. 373. 57-62 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] 賀佐 伸省: "新生化学実験講座" 東京化学同人, 4 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] 賀佐 伸省: "タンパク質化学" 廣川書店, 5 (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] S.Yonezawa: "Structural studies of rat cathepdin E: Aminoーterminal structure and carbohydrate units of mature enzyme." Biochem.Biophys.Res.Commun.166. 1032-1038 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] I.Ohkubo: "Human seminal plasma Znーα_2ーglycoprotein: Its purification and properties as compared with human plasma Znーα_2ーglycoprotein." Biochim.Biophys.Acta. 1034. 152-156 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] Y.Uehara: "Increased Nーacetylglucosamineー1ーphosphotransferase activity in sera from patients with leukemia." Jpn.J.Cancer Res.82. 82-85 (1991)

    • Related Report
      1990 Annual Research Report

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Published: 1990-04-01   Modified: 2016-04-21  

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