Identification of intracellular target molecules of glyco-genes and functional glycomics for those proteins

• Project/Area Number: 13670121
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: General medical chemistry
• Research Institution: Osaka University
• Principal Investigator: MIYOSHI Eiji Osaka University Graduate School of Medicine Div. Biochemistry, Associate Professor, 医学部, 助教授 (20322183)
• Project Period (FY): 2001 – 2002
• Project Status: Completed (Fiscal Year 2002)
• Budget Amount: ¥2,800,000 (Direct Cost: ¥2,800,000); Fiscal Year 2002: ¥600,000 (Direct Cost: ¥600,000); Fiscal Year 2001: ¥2,200,000 (Direct Cost: ¥2,200,000)
• Keywords: Hepatocellular carcinoma / Haptoglobin / two-dimensional electrophoresis / GnT-III / GnT-V / Matriptase / Cancer Metastasis / Glycomics / がん転移 / ハプトグロビン / 血管新生 / βカテニン

Research Abstract

I identified intracellular target proteins for GnT-III and GnT-V and analyzed functions of those proteins. When DEN was administered to GnT-III transgenic mice, tumor formation in the liver was dramatically suppressed as compared to control mice. However, a few number of GnT-III transgenic mice treated with DEN had tumors. When serum proteins of those mice were analyzed by two-dimensional ectrophoresis followed by lectin blot, 6 target proteins for GnT-III were identified. One of these proteins was haptoglobin beta- chain. Haptoglobin is a radical scavenger and its function in terms of aberrant glycosylation should be clarified (Free Radical Research 36, 327-833, 2002). I also identified matriptase as a target molecule for GnT-V. Matriptase is one of membrane-anchored serine-protease, which has recently cloned. When GnT-V was transfected into a human gastric cancer cell MKN45, experimental metastasis was markedly promoted. The mechanism was due to induction of matriptase secretion because matriptase bearing beta1-6 GlcNAc branching, a product of GnT-V had a prolongation for degradation. This is the first answer for a question why GnT-V promotes cancer metastasis.

Research Products

• [Publications] Kitada, T., Miyoshi, E., et al.: "Addition of bisecting N-acetylglucosamine residues to E-cadherin downregulates the tyrosine phosphorylation of β-catenin"J. Biol. Chem.. 276. 475-480 (2001)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Chakraborty A K, Miyoshi E, et al.: "Fusion-Hybrids with macrophage and melanoma cells up-regulate N-acetylglucosaminyltransferase V, β1-6 branching, and metastasis"Cell Growth and Differentiation. 12. 623-630 (2001)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Ekuni A, Miyoshi E et al.: "A glycomic approach to hepatic tumors in N-acetylglucosaminyltransferase III (GnT-III) transgenic mice induced by diethylnitorsamine (DEN) : Identification of haptoglobin as a target molecule of GnT-III"Free Radical Research. 36. 827-833 (2002)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Ihara S, Miyoshi E, et al.: "Prometastatic effect of N-acetylglucosaminyltransferase V is due to modification and stabilization of active matriptase by adding beta 1-6 GlcNAc branching"J. Biol. Chem.. 277. 16960-16967 (2002)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Saito T, Miyoshi E, et al.: "A secreted type of beta 1,6-N-acetylglucosaminyltransferase V (GnT-V) induces tumor angiogenesis without mediation of glycosylation. A novel function of GnT-V distinct from the original glycosyltransferase"J. Biol. Chem.. 277. 17002-17008 (2002)
  • Description: 「研究成果報告書概要(和文)」より
• [Publications] Kitada, T. Miyoshi. E., Noda, K., Higashiyama, S., Ihara, H., Matsuura, N., Hayashi, N., Kawata, S., Matsuzawa, Y., and Taniguchi N.: "Addition of bisecting N-acetylglucosamine residues to: E-cadherin downregulates the tyrosine phosphotylation of β-catenin"J. Biol. Chem.. 276. 475-480 (2001)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Chakraborty A K, Pawelek J, Ikeda Y, Miyoshi E, Kolesnikova N, Funasaka Y, Ichihashi M, and Taniguchi N.: "Fusion-Hybrids with macrophage and melanoma cells up-regulate N-acetylglucosaminyltransferase V, β1-6 branching, and metastasis"Cell Growth and Differentiation. 12. 623-630 (2001)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Ekuni A, Miyoshi E. Ko J H, Noda K, Kitada T, Ihara S, Endo T, Hino A, Honke K, and Taniguchi N: "A glycomic approach to hepatic tumors in N-acetylglucosaminyltransferase III (GnT-III) transgenic mice induced by diethylnitorsamine (DEN): Identification of haptoglobin as a target molecule of GnT-III"Free Radical Research. 36. 827-833 (2002)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Ihara S, Miyoshi E, Ko JH, Murata K, Nakahara S, Honke K, Dickspn RB, Lin CY, Taniguchi N: "Prometastatic effect of N-acetylglucosaminyltransferase V is due to modification and stabilization of active matriptase by adding β 1-6 GlcNAc branching"J. Biol. Chem.. 277. 16960-16967 (2002)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Saito T, Miyoshi E, Sasai K, Nakano N, Eguchi H, Honke K, Taniguchi N: "A secreted type of β 1,6-N-acetylglucosaminyltransferase V (GnT-V) induces tumor angiogenesis without mediation of glycosylation. A novel function of GnT-V distinct from the original glycosyltransferase"J. Biol. Chem.. 277. 17002-17008 (2002)
  • Description: 「研究成果報告書概要(欧文)」より
• [Publications] Kitada, T., Miyoshi, E., et al.: "Addition of bisecting N-acetylglucosamine residues to E-cadherin downregulates the tyrosine phosphorylation of β-catenin"J.Biol.Chem.. 276. 475-480 (2001)
• [Publications] Chakraborty A K, Miyoshi E, et al.: "Fusion-Hybrids with macrophage and melanoma cells up-regulate N-acetylglucosaminyltransferase V, β1-6 branching, and metastasis"Cell Growth and Differentiation. 12. 623-630 (2001)
• [Publications] Ekuni A, Miyoshi E et al.: "A glycomic approach to hepatic tumors in N-acetylglucosaminyltransferase III (GnT-III) transgenic mice induced by diethylnitorsamine (DEN) : Identification of haptoglobin as a target molecule of GnT-III"Free Radical Research. 36. 827-833 (2002)
• [Publications] Ihara S, Miyoshi E, et al.: "Prometastatic effect of N-acetylglucosaminyltransferase V is due to modification and stabilization of active matriptase by adding beta 1-6 GlcNAc branching"J.Biol.Chem.. 277. 16960-16967 (2002)
• [Publications] Saito T, Miyoshi E, et al.: "A secreted type of beta 1,6-N-acetylglucosaminyltransferase V (GnT-V) induces tumor angiogenesis without mediation of glycosylation. A novel function of GnT-V distinct from -the original glvcosvltransferase"J.Biol.Chem.. 277. 17002-17008 (2002)
• [Publications] T.Kitada, E.Miyoshi, N.Taniguchi et al.: "The addition of bisecting N-acetylglucosamine residues to E-cadherin downregulates the tyrosine phosphorylation of beta-catenin"J. Biol. Chem.. 276. 475-480 (2001)
• [Publications] W.Wang, E.Miyamoto, N.Taniguchi et al.: "Ectopic expression of α 1, 6 fucosyltransferase in mice causes steatosis in the liver and kidney accompanied by a modification of lysosomal acid lipase"Glycobiology. 11. 165-174 (2001)
• [Publications] N.Taniguchi, E.Miyoshi, et al.: "A glycomic approach to the identification and characterization of glycoprotein function in cells transfected with glycosyltransferase genes"Proteomics. 1. 239-247 (2001)
• [Publications] A.K.Chakraborth, E.Miyoshi, H.Taniguchi et al.: "Fusion Hybrids with Macrophage and Melanoma Cells Up-Regulate N-Acetylglucosaminyltransferase V, β 1-6 Branching, and Metastasis"Cell Growth Differ. 12. 623-630 (2001)
• [Publications] Y.Ito, E.Miyoshi, N.Taniguchi et al.: "Elevated expression of UDP-N-acetylglucosamine : alphamannoside beta 1, 6 N-acetylglucosaminyltransferase is an early event in hepatocarcinogenesis"Int. J. Cancer. 91. 631-637 (2001)
• [Publications] T.Saito, E.Miyoshi, N.Taniguchi et al.: "A secreted type of β 1, 6-N-acetylglucosaminyltransferase V (GnT-V)induced tumor angiogenesis without mediation of glycosylation;---A novel function of GnT-V distinct from the original glycosyltransferase activity---"J. Biol. Chem.. (in press). (2002)
• [Publications] A.Ekuni, E.Miyoshi, N.Taniguchi et al.: "A glycomic approach to hepatic tumors in N-acetylglucosaminyltransferase III (GnT-III) transgenic mice induced by diethylynitorsamine (DEN): Identification of haptoglogin as a target molecule of GnT-III"Free Rad. Res. (in press). (2002)
• [Publications] S.Ihara, E.Miyoshi, N.Taniguchi et al.: "Pro-metastatic effect of N-acetylgucosaminyltransferase V is due to modification and stabilization of active matriptase by adding β1-6 GlcNAc-branching"J. Biol. Chem.. (in press). (2002)
• [Publications] T.Saito, E.Miyoshi, N.Taniguchi et al.: "A secreted type of β 1, 6-N-acetylglucosaminyltransferase V (GnT-V)induced tumor angiogenesis without mediation of glycosylation;---A novel function of GnT-V distinct from the original glycosyltransferase activity---"J. Biol. Chem.. (in press). (2002)
• [Publications] N.Taniguchi, E.Miyoshi, et al.: "Glycans in Cell Interaction and Recognition Therapeutic Aspects"The involvement of bisecting N-Acetylglucosamine in cancer. 73-88 (2001)