| Project/Area Number |
02558014
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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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 | The University of Tokyo |
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Principal Investigator |
INOUE Yasuo The University of Tokyo, Faculty of Science, Associate Professor, 理学部, 助教授 (30004336)
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| Co-Investigator(Kenkyū-buntansha) |
KANAMORI Akiko University of Tokyo, Faculty of Science, JSPS Postdoc. Fellow, 理学部, 日本学術振興会特別研
YAMAGATA Tatsuya Mitsubishi Kasei Institute of Life Science, Director, 細胞認識研究部, 部長
KUDO Shigeharu Gunma University, School of Medicine, Lecturer, 医学部, 講師 (70008267)
INOUE Sadako Showa University, School of Pharmaceutical Sciences, Associate Professor, 薬学部, 助教授 (00053827)
KITAJIMA Ken The University of Tokyo, Faculty of Science, Senior Research fellow, 理学部, 助手 (80192558)
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| Project Period (FY) |
1990 – 1992
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| Project Status |
Completed (Fiscal Year 1992)
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| Budget Amount *help |
¥14,400,000 (Direct Cost: ¥14,400,000)
Fiscal Year 1992: ¥1,500,000 (Direct Cost: ¥1,500,000)
Fiscal Year 1991: ¥4,600,000 (Direct Cost: ¥4,600,000)
Fiscal Year 1990: ¥8,300,000 (Direct Cost: ¥8,300,000)
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| Keywords | Deaminated Neuraminic Acid / KDN / KDN-Glycans / KDN-Glycoproteins / KDN-Gangliosides / Anti-KDN Monoclonal Antibodies / CMP-KDN Synthetase / KDN-Transferases / KDN-糖脂質 / 抗KDN単クロ-ン抗体 / オリゴ・ポリKDN / KDNー糖タンパク質 / KDNー糖脂質 / オリゴKDN / 抗オリゴKDN抗体 |
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| Research Abstract |
In 1986 we first reported the natural occurrence of deaminated neuraminic acid (2-keto-3-deoxy-d-glycero-d-galactonononic acid, KDN) in fish egg glycoprotein. Since then, an increasing number of KDN-containing glycoconjugates have been reported. We suspected that development of more sensitive assay procedures might demonstrate KDN-glycoconjugates in tissues and cells of many additional animal types. The present investigation has, for its objective, the establishment of chemical and immunochemical probes for the detection of KDN-glycoconjugates.
(1) Detection and characterization of oligo/poly(sialic acid) and oligo/poly(KDN) units in glycoconjugates:- We developed new GLC and HPLC procedures for the identification-quantitation of nonulosonates and lower oligomers of NeuAc, NeuGc, and KDN. We examined and compared high-performance anion-exchange chromatographic separation of ^3H-labeled oligo(NeuAc), oligo(NeuGc), and oligo(KDN) alditols by using Mono-Q HR 5/5 resin. We also examined a m
… More
ethod of selective and quantitative microprecipitation for separation and purification of oligomers and polymers of polysialo- and poly(KDN)-glycoproteins.
(2) Generation and use of immunochemical probes for KDN-glycan chains:- The availability of KDN-glycoconjugates prompted us to generate monoclonal antibodies against the KDN-glycan chains found in KDN-ganglioside and KDN-glycoprotein. We first produced a monoclonal antibody IgG3 subclass (mAb.kdn3G) that specifically bound to (KDN)GM3, but not to (NeuAc)GM3. We also generated another monoclonal antibody IgM (mAb.kdn8kdn) by fusion of spleen cells from a BALB/c mouse immunized with KDN-gp and P3U1 mouse myeloma cells. The specificity of these monoclonal antibodies, mAb.kdn3G and mAb.kdn8kdn, was examined and the epitope structures were determined to be KDNalpha2*3Galbeta1* and KDNalpha2*8KDNalpha2* structures, respectively. The potential utility of these monoclonal antibodies is addressed in searching for KDN-glycoconjugates which contain these epitope structures. Less
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