1992 Fiscal Year Final Research Report Summary
Studies on the glycoproteins expressing neural specific carbohydrate
| Project/Area Number |
03680147
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| Research Category |
Grant-in-Aid for General Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Research Field |
物質生物化学
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| Research Institution | KYOTO UNIVERSITY |
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Principal Investigator |
KOZUTSUMI Yasunori Faculty of Pharmaceutical Sciences Kyoto Univ., Biological Chemistry Instructor, 薬学部, 助手 (70205425)
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| 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)
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| Project Period (FY) |
1991 – 1992
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| Keywords | HRP / Grasshopper / Neurons of insects / Carbohydrate chain |
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| 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.
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