Properties of the Endonuclease Secreted by Human B Lymphoblastic IM9 Cells

  • 발행 : 1998.01.31

초록

We have employed a DNA-native-polyacrylamide gel electrophoresis (DNA-native-PAGE) assay system to characterize the enzyme activity of the endonuclease secreted by human B lymphoblastic IM9 cells. Experimental results clearly demonstrated that the endonuclease activity of IM9 cell culture medium is distinct from that of DNase I in the DNA-native-PAGE assay system. Immunoprecipitation analysis using anti-DNase I antibody showed that the secreted endonuclease is not recognized by the antibody. The secreted endonuclease was estimated using supercoiled plasmid DNA as a substrate. The pH optimum required for the catalytic activity was determined to be in the range of pH 6.6-7.4. No significant difference in the endonuclease secretion was observed by stimulation of the IM9 cells with interferon-${\gamma}$ or interleukin-$1{\beta}$.

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참고문헌

  1. Adams, D. O. and Hamilton, T. A. (1984) The cell biology of macrophage activation. Annu. Rev. Immunol. 2. 289-318.
  2. Anzai, N., Kawabara, H., Hirama, T., Masutani, H., Ueda, Y., Yoshida, Y., and Okuma, M. (1995) Types of nuclear endonuclease activity capable of inducing internucleosomal DNA fragmentation are completely different between human $CD34^+$ cells and their granulocytic descendents. Blood 86, 917-923.
  3. BaIl, W. D. (1974) Isolation of salivary ribonuclease, deoxyribonuclease and amylase from the parotid gland of the rat, Rattus norvegicus albinus. Biochim. Biophys. Acta. 341, 305-317. https://doi.org/10.1016/0005-2744(74)90223-X
  4. Barry, M. A. and Eastman, A. (1993) Identification of deoxyribonuclease II as an endonuclease involved in apoptosis. Arch. Biochem. Biophys. 300, 440-450. https://doi.org/10.1006/abbi.1993.1060
  5. Chace, J. H., Abed, N. S., Adel, G. L., and Cowdery, J.S (1993) Regulation of differentiation in $CD5^+$ and conventional B cells. Sensitive to LPS-induced differentiation and interferon-gamma-mediated inhibition of differentiation. Clin. Immunol. Immunopathol. 68, 327-332. https://doi.org/10.1006/clin.1993.1134
  6. Cordis, G. A., Goldbaltt, P. J., and Deutscher, M. P. (1975) Purification and characterization of a major endonuclease from rat liver nuclei. Biochemistry 14, 2596-2603. https://doi.org/10.1021/bi00683a006