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| A Cytotoxic Ribonuclease Isolated from Occytes of Chinese Northeast Frog（Rana dybowskii） |
| TAO Feng-yun1,2,ZHAO Wei2,LIN Qiang2,MA Run-yu1 |
1.College of Life Science and Technology, Beijing University of Chemical Technology, Beijing 100029,China
2.Biochemical Engineering College, Beijing Union University , Beijing 100023,China |
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Abstract By acetone precipitation, SP-Trisacryl cation-exchange chromatography, Sephadex G-75 gel filtration chromatography and C8 reversed-phase chromatography, a single component ribonuclease is purified from oocytes of Chinese Northeast frog (Rana dybowskii). SDS-PAGE electrophoresis shows a single band with relative molecular weight of 13kDa. Its optimum temperature is 65℃, and optimum pH is 5.5~6.0. Its Michaelis constant is 4.11μmol/L, and maximum reaction rate is 2.82pmol/s. In vitro cytotoxicity experiments the IC50 value for HeLa，K562 and MCF-7 tumor cells is 0.6μmol/L, 0.8μmol/L and 4μmol/L separatedly, while for the normal human fibroblasts cells, even up to 8μmol/L, no cytotoxic effects were observed. This low molecular weight ribonuclease with selective cytotoxicity for tumor cells isolated from oocytes of the Chinese northeast frog(Rana Dybowskii) will join the candidate proteins of drug molecular for the treatment of cancer.
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Received: 07 January 2010
Published: 25 May 2010
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