2001 Fiscal Year Final Research Report Summary
Molecular Mechanisms Responsible for Glyco-chain Heterogeneity
| Project/Area Number |
11480173
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Structural biochemistry
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| Research Institution | Institute of Physical and Chemical Reserch |
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Principal Investigator |
SUZUKI Akemi Institute of Physical and Chemical Research, Group Director a Sphingolipid Expression Research Laboratory., スフィンゴ脂質発現制御研究チーム, グループディレクター(研究職) (70134533)
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| Co-Investigator(Kenkyū-buntansha) |
NAKAMURA Kyoko Institute of Physical and Chemical Research, Researcher, 糖鎖機能研究チーム, 研究員 (30124481)
HASIMOTO Yasuhiro Institute of Physical and Chemical Research, Laboratory Head, 糖鎖機能研究チーム, チームリーダー(研究職) (80164797)
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| Project Period (FY) |
1999 – 2001
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| Keywords | CMP- NeuAc hydroxyalse / tissue-specific regulation / glycibiology / diversity of glyco-chains |
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| Research Abstract |
The expression of N-glycolylneuraminic acid (NeuGc) is regulated by CMP-N-acetylneuraminic acid (CMP-NeuAc) hydroxylase in a rate limiting manner. The 92 bp deletion in human CMP-NeuAc hydroxylase gene is the cause of the lacking of NeuGc expression in human tissues. Brain tissues in mammals which can produce NeuGc in their visceral organs contain very low mount of NeuGc, and do not give detectable signals of CMP-NeuAc hydroxylase mRNA by Northern blottiug. These results suggest that brain has a molecular mechanism to suppress the transcription of the hydroxylase. We cloned a 13kb long DNA containing a 6.5 kb fragment flanking at 5' site of the hydroxylase gene. We sequenced it and constructed 6.5 kb and lkb fraglnents fused with luciferase cDNA as a reporter. L929 cells contained 28% NeuGc in total sialic acid and were transfected with above constructs.. The both constructs failed to detect transcription activity. We need to determine the start site of major transcript of L929 celis. Beta6GlcNAc transferase to transfer GlcNAcbetal-6 to GaINA of Galbetal-3GaINAc of glycolpids and glycoproteins is regulated in a kidney proximal tubular cell specific mauner. We took an advantage for using the inbred mice which are defective in this cell specific regulation as a negative control and identified a glycoprotein modified by beta6GlcNAc transferase. The glycoprotein was identified as megalin on the basis of its molecular weight and detection with antibody produced by a synthetic peptide of mouse megalin. We purified this protein and measured ligand binding activity using retual binding protein and fluorescence oorrelation spectrometry. Ligand binding activity of megalin was the same if megalin has different modification by glyco'cahins.
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Research Products
(12 results)
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[Publications] Sekine, M., Taya, C., Kikkawa, Y, Yonekawa, H., Takenka, M., Matsuoka, Y, Imai, E., Izawa, M., Kannagi, R., Suzuki, A: "Regulation of mouse kidney tubular epithelial cell-specific expression of core 2 GlcNAc transferase"Eur. J. Biochem.. 268. 1129-1135 (2001)
Description
「研究成果報告書概要(和文)」より
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[Publications] Suzuki, A., Omae, F., Enomoto, A., Yoshioka, S., Irie, A., Sekine, M., Kikkawa, Y., Taya.C., Yonekawa, H., Takenaka, M., Matsuoka, Y., Imai, E., Izawa, M., Kannagi, R: "Tissue specific control of glyco-chains"Int. Cong. Ser.. 1223. 29-37 (2001)
Description
「研究成果報告書概要(和文)」より
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[Publications] Kobayashi, T., Yokoyama, I., Suzuki, A., Abe, M., Hayashi, S., Matsuda, H., Morozumi, K., Breimer, M.E., Rydberg, L., Broth, C.G., Tibell, A., Korsgren, O., Takagi, H., Nakao, A.: "Lack of antibody production against Hanganutziu-Deicher(H-D)antigens with N-glycolylneuraminic acid in patients with porcine exposure histry. 7, 177-180 (2000)"Xenotransplantation. 7. 177-180 (2000)
Description
「研究成果報告書概要(和文)」より
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[Publications] Suzuki, A., Omae, F., Enomoto, A., Yoshio:ka, S., Me, A., Sekine, M., Kikkawa, Y., Taya. C., Yonekawa, H., Takenaka, M., Matsuoka, Y., Imai, E., Izawa, M., and Kannagi, R: "Tissue specific control of glyco-chains."Int. Cong. Ser. 1223. 29-37 (2001)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Uchida Y, Hara M., Nishio H., Sidransky E., Inoue S., Otsuka, E, Suzuki, A., Elias, P. M., Holleran, W. M., and Hamanaka S.: "Epidermal sphingiomyelins are precursors for selected stratum corneuml ceramide."J. Lipid Res.. 41. 2071-2081 (2000)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Sudo, T., Zhao, X., Wakamatsu, Y, Shibahara, M., Nomura, N., Nakahara, T., Suzuki, A., Kobayashi, Y, Jin, C., Murata, T., Yokoyama, K. K: "lnduction of the differentaition of human HL-60 promyeiocytic leukemia cell line by succinoyl trehalose lipids,"Cytotechnology. 33. 259-264 (2000)
Description
「研究成果報告書概要(欧文)」より
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[Publications] Kobayashi, T., Yokoyama, I., Suzuki A., Abe, M., Hayashi, S., Matsuda, H., Morozumi, K. Breimer, M. E., Rydberg, L., Broth, C. G., Tibell, A., Korsgren, O., Takagi, H., and Nakao, A: "Lack of antilbody production against Hanganutziu-Deicher (H-D)antigens with N-glyrolyrneuraminic acid in patients with porcine exposure histry."Xenotransplantation. 7. 177-180 (2000)
Description
「研究成果報告書概要(欧文)」より
