DNA Aggregating Protein from Carp (Cyprinus capio) Eggs(Biochemistry/Molecular Biology)

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概要

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• Hemagglutinating activity for glutaraldehyde-fixed, trypsinized human and rabbit erythrocytes was obtained from carp (Ciprinus capio) egg extract. This activity was inhibited by sulfated glycans such as heparin and fucoidan, and salmon testis DNA (stDNA), but not by mono- and oligosaccharides. The active fraction from hydroxyapatite column, HA400, caused aggregation of stDNA at a concentration of 125 μg/ml. HA400-induced aggregation was inhibited by heparin and fucoidan at the concentrations of 25 and 50 μg/ml, respectively. HA400 also bound to short size single- or double-stranded oligonucleotide in dot blot hybridization analysis using ^<32>P-labeled probe. HA400 was further purified by heparin-Sepharose column chromatography, and consequently, active fractions (HS2, HS3 and HS4) were obtained. Hemagglutinating activities of HS2 and HS4 were 16 and 32 times higher than that of HS3, respectively. In contrast, HS3 showed the highest DNA aggregating activity among these fractions, indicating that the activities of DNA aggregation and hemagglutination were separated by heparin-Sepharose column chromatography. HS3-induced stDNA aggregation was inhibited by 0.5 M NaCl or above. Southwestern blot analysis revealed that the DNA aggregating protein (DAP) possessing oligonucleotide-binding capacity was the 18kDa protein. The N-terminal amino acid sequence of DAP was LKEGE-CEVCVGFLGR having partial homology with DNA-binding protein HU-α, but showed no homology with nucleohistone proteins. These results indicate that there is a novel, non-histone DAP in carp eggs.

• 公益社団法人日本薬学会の論文
• 2005-01-01

著者

• TAKAYANAGI Motoaki

  Department of Pathophysiology, Tohoku Pharmaceutical University

• Ogawa Y

  Institute Of Molecular Biomembrane And Glycobiology Tohoku Pharmaceutical University

• Takayanagi M

  Department Of Pathophysiology Tohoku Pharmaceutical University

• Takayanagi Motoaki

  Department Of Pharmacology And Toxicology Cancer Research Institute Tohoku Pharmaceutical University

• Takayanagi Motoaki

  Cancer Research Institute Tohoku Pharmaceutical University

• Nitta K

  Institute Of Molecular Biomembrane And Glycobiology Tohoku Pharmaceutical University

• Nitta Kazuo

  Moa Health Science Foundation

• SUGAWARA Shigeki

  Institute of Molecular Biomembrane and Glycobiology, Tohoku Pharmaceutical University

• HOSONO Masahiro

  Institute of Molecular Biomembrane and Glycobiology, Tohoku Pharmaceutical University

• HOSONO Masahiro

  Cencer Research Institute, Tohoku Pharmaceutical University

• SUGAWARA Shigeki

  Cencer Research Institute, Tohoku Pharmaceutical University

• OGAWA Yukiko

  Cencer Research Institute, Tohoku Pharmaceutical University

• TAKAYANAGI Motoaki

  Cencer Research Institute, Tohoku Pharmaceutical University

• NITTA Kazuo

  Cencer Research Institute, Tohoku Pharmaceutical University

• Hosono Masahiro

  Institute Of Molecular Biomembrane And Glycobiology Tohoku Pharmaceutical University

• Sugawara Shigeki

  Institute Of Molecular Biomembrane And Glycobiology Tohoku Pharmaceutical University

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