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2001 Fiscal Year Final Research Report Summary

Study on the selection and traffic of lysosomal enzyme

Research Project

Project/Area Number 12680704
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Cell biology
Research InstitutionTokyo University of Agriculture and Technology (2001)
Okayama University of Science (2000)

Principal Investigator

NISHIKAWA Atsushi  okyo University of Agriculture and Technology, Dep. Of Applied Bio logical Science Associate Prof., 農学部, 助教授 (30218127)

Project Period (FY) 2000 – 2001
KeywordsIysosome / N-linked / Dnase I / manuose-6-phosphate / Oligosacchary Itames ferase
Research Abstract

In many cell types, newly synthesized acid hydrolases acquire mannose 6-phosphate residues which serve as high affinity ligands for binding to Man-6-P receptors and targeting to lysosomes. A key step in this pathway is the ability of the enzyme UDP-GlcNAc : Lysosomal enzyme N-acetylglucosaminyl phosphotransferase to selective recognize and phosphorylate acid hydrolases. We have previously demonstrated that phosphotransferase recognizes conformational dependent protein determinants present on the acid hydrolases and absent in most nonlysosomal glycoproteins. It was recently reported that the secretory glycoprotein DNase I contains Man-6-P residues and we found that the human, bovine and mouse forms of this enzymes are phosphorylated to a low extent when expressed in COS cells. The poor phosphorylation of human and bovine DNase I is due to the lack of two critical lysines (position 27 and 74) present in the mouse enzyme whereas the poor phosphorylation of mouse Dnase I appears to be due to the presence of "inhibitory" amino acid (Val23) that block the phosphotransferase recognition domain and the disappear of two critical amino acids (Tyr54 and Serl90).
Then, we also report the substrate specificity of GPT for the oligosaccharides using variety of fluorescence labeled high mannose type oligosaccharides. As a point to be noticed, GPT can transfer the GlcNAc-phosphate to the GlcMan9GlcNAc2 that still remains glucose.

  • Research Products

    (12 results)

All Other

All Publications (12 results)

  • [Publications] H.Yamamoto, J.Swoger, S.Greene, T.Saito, A.Nishikawa, J.R.Moskal他8名: "b1,6-N-Acetylglucosamine-bearing N-glycans in human gliomas : Implications for a role in regulating invasivity"Cancer Res.. 60. 134-142 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] A.Nishikawa, S.Mizuno: "The efficiency of N-linked glycosylation of bovine DNase I depends on the Asn-X-Ser/Thr sequence and the tissue of origin"Biochem. J. 354. 245-248 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] N.Taniguchi, A.Ekuni, E.Miyoshi, Y.Ikeda, Y.Ihara, A.Nishikawa, K.Honke, Takahashi, M.: "A glycomic approach to the identification and characterization of glycoprotein function in cells transfected with glycosyltransferase genes"Proteomics. 1. 239-247 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] A.Nishikawa, C.Kanehira, K.Imamura: "Purification and characterization of pyruvate kinase from lamprey (Entosphenus japonicus) muscle"Comp. Biochem. Phys. B,. 128. 157-162 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] A.Nishikawa: "Study on the lysosomal enzyme selection system"J. Appl. Glycosci.. 49(in press). (2002)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] A.Nishikawa: "Lysosomal enzyme GlcNAc-1-phosphotransterase pp611-616 (Book title : Handbook of glycosyltransferases and related genes)"N. Taniguchi, K. Honke and M. Fukuda, Eds. Springer. 670 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] H.Yamamoto, J. Swoger, S. Greene,T. Saito,J. Hurh, C. Sweeley, J. Leestma, E. krdichian, L. Cerullo, A. Nishikawa, Y. Ihara, N. Taniguchi, J. R. MoskaI: "β1, 6-N-Acetylglucosamine-bearing N-glycans in human gliomas : Implications for a role in regulating invasivity"Cnacer Res.. 60. 134-142 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] N.Taniguchi,A. Ekuni, E. Miyoshi.Y. Ikeda,Y, Ihara, A. Nishikawa, K. Honke, Takahashi, M.: "A glycomic approach to the identification and characterization of glycoprotein function in cells transfected with glycosyltransferase genes"Biochem. J.. 355. 245-248 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] A. Nishikawa, C. Kanehira and K. Imamura: "Purification and characterization of pyruvate kinase from lamprey (Entosphenus japonicus) muscle"Proteomics. 1. 239-247 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] A. Nishikawa, S. Mizuno: "The efficiency of N-linked glycosylation of bovine DNase I depends on the Asn-X-Ser/Thr sequence and the tissue of origin"Comp. Blochem. Phys. B.. 128. 157-162 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] A. Nishikawa: "Study on the lysosomal enzyme selection system"J. Appl. Glycosci. 49(in press). (2002)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] A. Nishikawa: "Lysosomal enzyme GlcNAc-l -phosphoransferase"N.Taniguchi, K. Honke and M. Fukuda Eds., Handbook of glycosyltransferases and related genes, pp611-616, Springer. (2001)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2003-09-17  

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