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A new glycome analysis by changing carbohydrate-binding specificities of cargo receptors in the cells

Research Project

Project/Area Number 13470485
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field Biological pharmacy
Research InstitutionThe University of Tokyo

Principal Investigator

YAMAMOTO Kazuo  The University of Tokyo, Graduate School of Frontier Sciences, Professor, 大学院・新領域創成科学研究科, 教授 (20174782)

Co-Investigator(Kenkyū-buntansha) MATSUMOTO Naoki  The University of Tokyo, Graduate School of Frontier Sciences, Associate Professor, 大学院・新領域創成科学研究科, 助教授 (40239108)
Project Period (FY) 2001 – 2003
Project Status Completed (Fiscal Year 2003)
Budget Amount *help
¥14,400,000 (Direct Cost: ¥14,400,000)
Fiscal Year 2003: ¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2002: ¥4,900,000 (Direct Cost: ¥4,900,000)
Fiscal Year 2001: ¥6,000,000 (Direct Cost: ¥6,000,000)
Keywordscargo receptor / ERGIC-53 / VIP36 / lectin / sugar specificity
Research Abstract

In secretory pathway of eucaryotic cells, newly synthesized glycoproteins are transported from the ER through the Golgi toward cell surface via transport vesicles. During this process, sorting events are performed by cargo receptors. ERGIC-53 and VIP36 having carbohydrate-binding domain homologous to leguminous lectins at lumenal region are receptors to direct specific proteins into vesicles. cDNA coding lectin domains of both receptors were constructed and expressed in E.coli. Interactions of these recombinant soluble ERGIC-53 and VIP36 with glycoproteins were analyzed using surface plasmon resonance with or without calcium ions under several pH conditions. Interaction of ERGIC-53 and IgY was not so different under the pH ranging from 6.5 to 7.2. By contrast, soluble VIP36 bound to porcine thyroglobulin stronger under the acidic condition than neutral pH. Oligomerization of soluble VIP36 occurred in relation to the enhanced association with the ligands under acidic condition. Then random oligonucleotides were introduced into cDNAs in place of the nucleotides encoding carbohydrate-binding loops of VP36 and ERGIC-53, respectively. The mutated cDNAs were expressed in CHO cells and the cells having different sugar moieties on the surfaces were enriched by flow cytometry. After cloning of the enriched cells, which have specifically changed to bind to PNA after the trasfection of mutated VIP36 cDNAs, 13 kinds of stable clones were established. These clones were classified into two groups based on the mutated cDNAs expressed in the cells and the carbohydrate-binding loops of these mutated VIP36 cDNAs were deduced to be DPDSNGGSF and DSSFNYVHA, respectively.

Report

(4 results)
  • 2003 Annual Research Report   Final Research Report Summary
  • 2002 Annual Research Report
  • 2001 Annual Research Report
  • Research Products

    (23 results)

All Other

All Publications (23 results)

  • [Publications] N.Higashi: "The macrophage C-type lectin specific for galactose/N-acetylgalactosamine is an endocytic receptor expressed on monocyte-derived immature dendritic cells."J.Biol.Chem.. 277. 20686-20693 (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] M.Mitsuki: "A species-specific determinant on b2-microglobulin required for Ly49A recognition of its MHC class I ligand."Int.Immunol.. 16. 197-204 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] H.Wada: "The inhibitory NK cell receptor CD94/NKG2A and the activating receptor CD94/NKG2C bind the top of HLA-E through mostly shared but partly distinct sets of HLA-E residues."Eur.J.Immunol.. 34. 81-90 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] K.Tajima: "Augmentation of NK cell-mediated cytotoxicity to tumor cells by inhibitory NK cell receptor blockers"Int.Immunol.. 16. 385-393 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] 山本一夫: "細胞内輸送・選別における糖鎖認識プロセス"生化学. 76. 240-255 (2004)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] N.Higashi, K, Fujioka Denda-Nagai K, Hashimoto S, Nagai S, Sato T, Fujita Y, Morikawa A, Tsuiji M, Miyata-Takeuchi M, Sano Y, Suzuki N, Yamamoto K, Matsushima K, Irimura T.: "The macrophage C-type lectin specific for galactose N-acetylgalactosamine is an endocytic receptor expressed on monocyte-derived immature dendritic cells."J.Biol.Chem.. 277. 20686-20693 (2002)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] M.Mitsuki, N.Matsumoto, K.Yamamoto.: "A species-specific determinant on b2-microglobulin required for Ly49A recognition of its MI-IC class I ligand."Int.Immunol.. 16. 197-204 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] H.wada, N.Matsumoto, K.Maenaka, K.Suzuki, K.Yamamoto.: "The inhibitory NK cell receptor CD94/NKG2A and the activating receptor CD94/NKG2C bind the top of HLA-E through mostly shared but partly distinct sets of HLA-F residues."Eur.J.Immunol.. 34. 81-90 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] K.Tajima, N.Matsumoto, K.Ohmori, H.Wada, K.Suzuki, K.Yamamoto.: "Augmentation of NK cell-mediated cytotoxicity to tumor cells by inhibitory NK cell receptor blockers"Int.mmunol.. 16. 385-393 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] K.Yamamoto.: "Intracellular transport and sorting of glycoproteins by cargo receptors."Seikagaku. 76. 240-255 (2004)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2003 Final Research Report Summary
  • [Publications] M.Mitsuki: "A species-specific determinant on b2-microglobulin required for Ly49A recognition of its MHC class I ligand."Int.Immunol.. 16・2. 197-204 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] H.Wada: "The inhibitory NK cell receptor CD94/NKG2A and the activating receptor CD94/NKG2C bind the top of HLA-E through mostly shared but partly distinct sets of HLA-E residues."Eur.J.Immunol.. 34・2. 81-90 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] K.Tajima: "Augmentation of NK cell-mediated cytotoxicity to tumor cells by inhibitory NK cell receptor blockers."Int.Immunol.. 16・3. 385-393 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] K.Yamamoto: "細胞内輸送・選別における糖鎖認識プロセス"生化学. 76・3. 240-255 (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Sakamoto H.: "Modulation of heat-shock protein 27 (Hsp27) anti-apoptotic activity by methylglyoxal modification"J. Biol. Chem.. 277. 45770-45775 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Sun Y.: "Characterization and expression of L-amino acid oxidase of mouse milk"J. Biol. Chem.. 277. 19080-19086 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] Higashi N.: "The macrophage C-type lectin specific for galactose/N-acetylgalactosamine is an endocytic receptor expressed on monocyte-derived immature dendritic cells"J. Biol. Chem.. 277. 20686-20693 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] K.Yamamoto: "Protein Protocols Handbook, 2nd Edition"Affinity chromatography of oligosaccharides and glycopeptides with immobilized lectins. 917-931 (2002)

    • Related Report
      2002 Annual Research Report
  • [Publications] N.Matsumoto: "The functional binding site for the c-type lectin-line NK cell receptor Ly49A spans three domains of its MHC class I ligand"J.Exp.Med.. 193. 147-157 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] N.Matsumoto: "The c-type lectin-like NK cell receptor Ly49A recognizes furface of MHC class I ligand composed of three domains"Int.Immunol.. 13(5). 615-623 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] N.Matsumoto: "The NK cell MHC class I receptor Ly49A detects mutations on H-2Dd inside and outside of the peptide binding groove"J.Immunol.. 166. 4422-4428 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] R.Tozawa: "Asialoglycoprotein receptor deficiency in mice lacking the major receptor subunit : Obligate requirement of HL-1 for the stable expression of oligomeric receptor"J.Biol.Chem.. 276. 12624-12628 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] K.Yamamoto: "Protein Protocols Handbook, 2nd Edition(Affinity chromatography of oligosaccharides and glycopeptides with immobilized lectins"917-931 (2002)

    • Related Report
      2001 Annual Research Report

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Published: 2001-04-01   Modified: 2016-04-21  

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