Nucleic Acids Research, 2000, Vol. 28, No. 21 4097-4104
© 2000 Oxford University Press
Purification and cloning of cytotoxic ribonucleases from Rana catesbeiana (bullfrog)
1Institute of Biomedical Sciences, Academia Sinica, Taipei 115, Taiwan, 2Institute of Biochemistry, College of Medicine, National Taiwan University, Taipei 100, Taiwan, 3Department of Life Sciences, National Tsing-Hua University, Hsin-Chu 300, Taiwan and 4Graduate Institute of Life Sciences, National Defense Medical Center, Taipei 114, Taiwan
Ribonucleases with antitumor activity are mainly found in the oocytes and embryos of frogs, but the role of these ribonucleases in frog development is not clear. Moreover, most frog ribonuclease genes have not been cloned and characterized. In the present study, a group of ribonucleases were isolated from Rana catesbeiana (bullfrog). These ribonucleases in mature oocytes, namely RC-RNase, RC-RNase 2, RC-RNase 3, RC-RNase 4, RC-RNase 5 and RC-RNase 6, as well as liver-specific ribonuclease RC-RNase L1, were purified by column chromatographs and detected by zymogram assay and western blotting. Characterization of these purified ribonucleases revealed that they were highly conserved in amino acid sequence and had a pyroglutamate residue at their N-termini, but possessed different specific activities, base specificities and optimal pH values for their activities. These ribonucleases were cytotoxic to cervical carcinoma HeLa cells, but their cytotoxicities were not closely correlated to their enzymatic specific activities. Some other amino acid residues in addition to their catalytic residues were implicated to be involved in the cytotoxicity of the frog ribonucleases to tumor cells. Because the coding regions lack introns, the ribonuclease genes were cloned by PCR using genomic DNA as template. Their DNA sequences and amino acid sequences are homologous to those of mammalian ribonuclease superfamily, 
50 and 25%, respectively.
* To whom correspondence should be addressed. Tel: +886 2 2789 9167; Fax: +886 2 2782 9142; Email: ydliao@ibms.sinica.edu.tw +AF039104, AF242553–AF242556, AF288642
CiteULike 
Connotea 
Del.icio.us What's this?
This article has been cited by other articles:
|
|
 |
|
  M.-a. I. Economopoulou, E. G. Fragoulis, and D. C. Sideris Molecular cloning and characterization of the human RNase {kappa}, an ortholog of Cc RNase Nucleic Acids Res., October 8, 2007; 35(19): 6389 - 6398. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y.-C. Huang, Y.-M. Lin, T.-W. Chang, S.-J. Wu, Y.-S. Lee, M. D.-T. Chang, C. Chen, S.-H. Wu, and Y.-D. Liao The Flexible and Clustered Lysine Residues of Human Ribonuclease 7 Are Critical for Membrane Permeability and Antimicrobial Activity J. Biol. Chem., February 16, 2007; 282(7): 4626 - 4633. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
T. Nitto, K. D. Dyer, M. Czapiga, and H. F. Rosenberg Evolution and Function of Leukocyte RNase A Ribonucleases of the Avian Species, Gallus gallus J. Biol. Chem., September 1, 2006; 281(35): 25622 - 25634. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 Y.-D. Liao, J.-C. Jeng, C.-F. Wang, S.-C. Wang, and S.-T. Chang Removal of N-terminal methionine from recombinant proteins by engineered E. coli methionine aminopeptidase Protein Sci., July 1, 2004; 13(7): 1802 - 1810. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y.-D. Liao, S.-C. Wang, Y.-J. Leu, C.-F. Wang, S.-T. Chang, Y.-T. Hong, Y.-R. Pan, and C. Chen The structural integrity exerted by N-terminal pyroglutamate is crucial for the cytotoxicity of frog ribonuclease from Rana pipiens Nucleic Acids Res., September 15, 2003; 31(18): 5247 - 5255. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y.-J. Leu, S.-S. Chern, S.-C. Wang, Y.-Y. Hsiao, I. Amiraslanov, Y.-C. Liaw, and Y.-D. Liao Residues Involved in the Catalysis, Base Specificity, and Cytotoxicity of Ribonuclease from Rana catesbeiana Based upon Mutagenesis and X-ray Crystallography J. Biol. Chem., February 21, 2003; 278(9): 7300 - 7309. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Zhang, K. D. Dyer, and H. F. Rosenberg Human RNase 7: a new cationic ribonuclease of the RNase A superfamily Nucleic Acids Res., January 15, 2003; 31(2): 602 - 607. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P.-C. Tang, H.-C. Huang, S.-C. Wang, J.-C. Jeng, and Y.-D. Liao Regulation of ribonuclease expression by estradiol in Rana catesbeiana (Bullfrog) Nucleic Acids Res., July 15, 2002; 30(14): 3286 - 3293. [Abstract] [Full Text] [PDF]
|
|