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Studies on the glycoproteins expressing neural specific carbohydrate

Research Project

Project/Area Number 03680147
Research Category

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

Allocation TypeSingle-year Grants
Research Field 物質生物化学
Research InstitutionKYOTO UNIVERSITY

Principal Investigator

KOZUTSUMI Yasunori  Faculty of Pharmaceutical Sciences Kyoto Univ., Biological Chemistry Instructor, 薬学部, 助手 (70205425)

Co-Investigator(Kenkyū-buntansha) OKA S  Kyoto Univ. Biological Chem., Inst., 薬学部, 助手 (60233300)
KAWASAKI T  Kyoto Univ. Biological Chem., Prof., 薬学部, 教授 (50025706)
Project Period (FY) 1991 – 1992
Project Status Completed (Fiscal Year 1992)
Budget Amount *help
¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1992: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1991: ¥1,200,000 (Direct Cost: ¥1,200,000)
KeywordsHRP / Grasshopper / Neurons of insects / Carbohydrate chain / ピリジルアミノ化 / 昆虫 / 神経細胞 / 西洋ワサビペルオキシダ-ゼ(HRP) / マンノ-ス / コンカナバリンA / アフィニティ-カラム
Research Abstract

With progress in immunological techniques and histochemical methods, neuron-specific carbohydrate moieties have been discovered. These glycoconjugates carrying the neural specific carbohydrates are mainly observed on the cell surface and expressed in region- or stage-specific manner during the development of the nervous system. Therefore, these neuron- specific carbohydrate moieties may play important roles in recognition and/or migration of neural cells. Antiserum raised against horseradish peroxidase(HRP)specifically labels the nervous system of grasshopper and other insects and has been shown to recognize carbohydrate moiety that is expressed on the surface of neurons.In order to determine the structure of the carbohydrate epitope, we isolated the glycoprotein which express the neuron- specific carbohydrate antigen, from grasshopper nervous system using lectin chromatography. The glycoprotein showed apparent molecular weight of 35 kDa on SDS-PAGE under reducing condition. Removal of alpha-linked mannose residues by digestion with alpha-mannosidase completely destroyed the binding activity of the 35 kDa glycoprotein to the anti-HRP antibody. The carbohydrate composition of the 35 kDa glycoprotein was analyzed using a HPLC system with a pulsed amperometry detector. The 35 kDa glycoprotein was composed of Fuc, Man, Gal, Glc, GalNH2, and GlcNH2 with a molecular ratio of 1.0,1.4, 0.4, 1.5, 0.4, and 1.5, respectively.

Report

(3 results)
  • 1992 Annual Research Report   Final Research Report Summary
  • 1991 Annual Research Report
  • Research Products

    (7 results)

All Other

All Publications (7 results)

  • [Publications] A.KUROSAKA et al.: "The structure of a neural specific carbohydrate epitope of horseradish peroxidase recognized by anti-horseradish peroxidase antiserum." J.Biol.Chem.266(7). 4168-4172 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 黒坂 光,岡 昌吾,脇本 公嗣,川嵜 敏祐: "昆虫神経細胞に特異的に発現する糖鎖抗原" 蛋白質核酸酵素. 37(11). 1920-1923 (1992)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Y.KOZUTSUMI et al.: "Recognition of CMP-N-acetylneuraminic acid hydroxylation activity using a mouse liver cytosol fraction and soluble cytochrome 65purified from horce er" J.Biochem.110. 429-435 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] A. Kurosaka et al.: "The structure of a neural Specific Carbohydrate epitope of horseradish peroxidase recognized by anti-horseradish peroxidase antiserum" J. Biol. Chem.266 (7). 4168-4172 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] Y. Kozutsumi et al.: "Reconstitution of CMP-N-acetylneuraminic acid hydroxylation activity using a mouse liver cytosol fraction and soluble cytochrome b5 purified from horse erythrocytes" J. Biochem.110 (3). 429-435 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1992 Final Research Report Summary
  • [Publications] 黒坂 光 岡 昌吾 脇本 公嗣 川嵜 敏祐: "昆虫神経細胞に特異的に発現する糖鎖抗原" 蛋白質核酸酵素. 37(11). 1920-1923 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] Akira Kurosaka: "The Structure of a Neural Specific Carbohydrate Epltope of Horseradish Peroxidase Recogniged by Anti‐horseradish peroxidase Antiserum" The Journal of Biological Chemistry. 266. 4168-4172 (1991)

    • Related Report
      1991 Annual Research Report

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

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