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1997 Fiscal Year Final Research Report Summary

ANALYSIS OF ADP-RIBOSYLATION ON MOUSE LYMPHOCYTES USING BIOTINYL NAD

Research Project

Project/Area Number 08680687
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Functional biochemistry
Research InstitutionSHIMANE MEDICAL UNIVERSITY

Principal Investigator

TSUCHIYA Mikako  SHIMANE MEDICAL UNIV.BIOCHEMISTRY.PROFESSOR, 医学部, 教授 (90188582)

Project Period (FY) 1996 – 1997
KeywordsADP-ribosyltransferase / ADP-ribosylation / biotinyl NAD / lymphocytes
Research Abstract

Biotinyl NAD was prepared from 5- (N-succinimidyloxycarbonyl) pentyl D-biotinamide and N6 [[(6-aminohexyl) carbamoyl] methyl] NAD,purified by HPLC and confirmed to be a substrate for ADP-ribosylation catalyzed by chicken and mouse ADP-ribosyltransferases. When biotinyl NAD was used as a substrate for detection of specific AD-ribosylation of tuftsin in chicken plasma, the modification of the peptide was not detected by biotin-avidin-peroxydase reaction. After incubated with biotinyl NAD,mouse lymphocytes were stained with FITC-avidin and analyzed by cell sorter. The intensity of the fluorescent labeling of the cells did not decrease by addition of ADP-ribose, NAD or arginine in the labeling reaction or ADP-ribosylarginine hydrolase treatment after the labeling. The cells which do not have the ADP-ribosyltransferase were also stianed to a similar extent. Thus, the labeling does not exactly reflect the ADP-reibosylation occurring on the lymphocytes under the conditions we used for staining and washing of cells, probably because of non-specific and non-covalent binding of biotinyl NAD.To determine the subset of lymphocytes which have ADP-ribosyltransferase of its target protein, more stringent conditions may bed needed for labeling and washing.

  • Research Products

    (16 results)

All Other

All Publications (16 results)

  • [Publications] Nobumasa Hara: "Glutamic acid 207 in rodent T-cell RT6 antigens is essential for arginine-specific ADP-ribosylation" J.Biol.Chem.271(47). 29552-29555 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tsuyoshi Miyaoka: "Activation of Clostridium botulinum C3 exoenzyme-catalyzed ADP-ribosylation of RhoA by K^+ in a Mg2^+-dependent manner" J.Biochem.119(1). 200-207 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Masaharu Terashima: "Exocytosis of arginine-specific ADP-ribosyltransferase and p33 induced by A23187 and calcium or serum-opsonized zymosan in chicken polymorphonuclear leukocytes" J.Biochem.120(6). 1209-1215 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Tsuyoshi Miyaoka: "Immunohistochemical localization of ADP-ribosylarginine hydrolase in rodent CNS." Brain Res.746. 1-9 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Motoi Itoga: "Nitric oxide-induced modification of glyceraldehyde-3-phosphate dehydrogenase with NAD^+ is not ADP-ribosylation." J.Biochem.412. 227-232 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Masaharu Terashima: "ADP-ribosylation of tuftsin suppresses its receptor-binding capacity and phagocytosis-stimulating activity to murine peritoneal macrophages." FEBS Lett.412. 227-232 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Mikako Tsuchiya: "ADP-ribosylation in animal tissue" Plenum Press, 4 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Makoto Shimoyama: "ADP-ribosylation in animal tissue" Plenum Press, 8 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Nobumasa Hara: "Glutamic acid 207 in rodent T-cell RT6 antigens is essential for arginine-specific ADP-ribosylation." J.Biol.Chem.271 (47). 29552-29555 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tsuyoshi Miyaoka: "Activation of clostridium botulinum C3 exoenzyme-catalyzed ADP-ribosylation of RhoA by K+ in a Mg2+dependent manner." J.Biochem.119 (1). 200-207 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Masaharu Terashima: "Exocytosis of arginine-specific ADP-ribosyltransferase and p33 induced by A23187 and calcium or serum-opsonized zymosan in chicken polymorphonuclear leukocytes" J.Biochem.120 (6). 1209-1215 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Tsuyoshi Miyaoka: "Immunohistochemical localization of ADP-ribosylarginine hydrolase in rodent CNS." Brain Res. 746. 1-9 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Motoi Itoga: "Nitric oxide-induced modification of glyceraldehyde-3-phosphate dehydrogenase with NAD+ is not ADP-ribosylation." J.Biochem.121 (6). 1041-1046 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Masaharu Terashima: "ADP-ribosylation of tuftsin suppresses its receptor-binding capacity and phagocytosis-stimulating activity to murine perioneal macrophages." FEBS Lett.412. 227-232 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Mikako Tsuchiya: A newly identified glycosylphosphatidylinositol-anchored arginine-specific ADP-ribosyltransferase in chiken spleen ADP-ribosylation in animal tissue.Plenum Press, 245-248 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Makoto Shimoyama: Molecular cloning and characterization of arginine-specific ADP-ribosyltransferases from chicken bonne marrow cells. Plenum Press, 137-144 (1997)

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

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Published: 1999-03-16  

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