2007 Fiscal Year Final Research Report Summary
Biosynthesis and biological roles of specific sulfation pattern of chondroitin sulfate
| Project/Area Number |
18570125
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Functional biochemistry
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| Research Institution | Aichi University of Education |
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Principal Investigator |
HABUCHI Osami Aichi University of Education, Education, Professor (90024067)
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| Project Period (FY) |
2006 – 2007
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| Keywords | chondroitin sulfate / sulfotransferases / gene knockout / chondroitin sulfate E / C6ST / GalNAc4S-6ST / N-glycan / bone marow derived mast cells |
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| Research Abstract |
1. We generated GalNAc4S-6ST KO mice. GalNAc4S-65T-/- mice lost synthetic activity of CS-E completely in every tissues. Bone marrow derived mast cells derived from the GalNAc4S-6ST-/- mice could not synthesize CS-E and showed decreased activity of tryptase.
2. We established stable clone expressing GalNAc4S-6ST from rat chondrosarcoma cell line. This clone synthesized and secreted proteoglycan containing CS-E. Chain length of CS produced by the clone was much shorter than the length of CS produced by the parent cells, suggesting that formation of GalNAc (4,6-SO4) residues by GalNAc4S-6ST may inhibit chain elongation.
3. We established a new method to synthesize a unique nonrreducing terminal sequence GalNAc(4,6-SO_4)-GlcA(2SO_4)-GalNAc(6SO_4), using 2OST and GalNAc4S-6ST from GalNAc(4SO_4)-GlcA-GalNAc(6SO_4).
4.We synthesized various C6ST-1 mutants devoid of N-glycosylation sites by the site-directed mutagenesis, and showed that N-glycans attached to C6ST-1 contributed to production of active enzyme, intracellular localization and substrate specificity of C6ST-I.
5.We cloned squid GalNAc4S-63T cDNA from the squid cartilage and prepared the recombinant enzyme. CS-E synthesized by the recombinant GalNAc4S-6ST inhibit thrombin in the antithrombin III-independent manner
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[Journal Article] Expression of sulfotransferases involved in the biosynthesis of chondroitin sulfate E in the bone marrow derived mast cells2008
Author(s)
Ohtake, S., Kondo, S., Morisaki, T., Matsumura, K., Kimata, K., and Habuchi, O.
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Journal Title
Biochim. Biophys. Acta 1780
Pages: 687-695
Description
「研究成果報告書概要(和文)」より
Peer Reviewed
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[Journal Article] Expression of sulfotransferases involved in the biosynthesis of chondroitin sulfate E in the bone marrow derived mast cells2008
Author(s)
Ohtake, S., Kondo, S., Morisaki, T., Matsumura, K., Kimata, K, Habuchi, O
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Journal Title
Biochim. Biophys. Ada 1780
Pages: 687-695
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] The utility of formalin-fixed paraffin-embedded tissue blocks for quantitative analysis of N-acetylgalactosamine 4-sulfate 6-O-sulfotransferase mRNA expressed by colorectal cancer cells2007
Author(s)
Ito, Y., Watanabe, M., Nishizawa, T., Omachi, T., Kobayashi, T., Habuchi, O., and Nakayama
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Journal Title
Acta Histochem. Cytochem 40
Pages: 53-59
Description
「研究成果報告書概要(和文)」より
Peer Reviewed
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[Journal Article] The utility of formalin-fixed paraffm-embedded tissue blocks for quantitative analysis of N-acetylgalactosamine 4-sulfate 6-0-sulfotransferase mRNA expressed by colorectal cancer cells2007
Author(s)
Ito, Y, Watanabe, M., Nishizawa, T, Omachi, T., Kobayashi, T., Habuchi, O., Nakayama
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Journal Title
Acta Histochem. Cytochem 40
Pages: 53-59
Description
「研究成果報告書概要(欧文)」より
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[Journal Article] Determination of Substrate Specificity of Sulfotransferases and Glycosyltransferases (Proteoglycans)2006
Author(s)
Habuchi, H., Habuchi, O., Uchimura, K., Kimata. K, Muramatsu, T
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Journal Title
Methods Enzymo 416,5,14
Pages: 225-243
Description
「研究成果報告書概要(欧文)」より
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[Book] Knockout Mice and Proteoglycans in COMPREHENSIVE GLYCOSCIENCE.(Kamerling, J. P., et. al.eds.)Vol.3, Chapter 4.10, 159-1912007
Author(s)
Kimata, K., Habuchi, O., Habuchi, H., and Watanabe, H
Total Pages
32
Publisher
Elsevier
Description
「研究成果報告書概要(和文)」より
