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High Expression in E. coli and Biological Activity of T-19 Peptide
Corresponding Author(s) : Shujing Wang
Asian Journal of Chemistry,
Vol. 26 No. 11 (2014): Vol 26 Issue 11
Abstract
To obtain antitumor peptide of Tumstatin (T-19 peptide) and study its biological activity are reported in this paper. According to the nucleotide sequence of T-19 peptide and E. coli favorite to base, the gene of T-19 was synthesized and inserted into the vector pTYB2.After the recombined vector T-19-pTYB2 was transformed into BL-21 (DE3) E. coli competent cells. Transformed E. coli BL-21DE3 were induced by isopropyl-b-thiogalactopyranoside (IPTG) and high expressed under the condition 28 ºC, 0.1 mmol/L IPTG, 6 h. By 1,4-dithiothreitol (DTT) induced, T-19 peptide was obtained from chitin affinity chromatograph. The rate of its protein purification was as much as 90.83 % by high performance liquid chromatography. Its biological activity was identified by HE stain, TUNEL assay, transmission electron microscopic (TEM). T-19 peptiade can inhibit human hela cell proliferation,promote human hela cell apoptosis and have no affect on endothelial cell. It would be an efficient method for tumor therapy in future, which will lay foundation on its mechanism of action research and clinical tumor therapy.
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References
J. Saus, J. Wieslander, J.P. Langeveld, S. Quinones and B.G. Hudson, J. Biol. Chem., 263, 13374 (1988).
Y. Maeshima, P.C. Colorado, A. Torre, K.A. Holthaus, J.A. Grunkemeyer, M.B. Ericksen, H. Hopfer, Y. Xiao, I.E. Stillman and R. Kalluri, J. Biol. Chem., 275, 21340 (2000); doi:10.1074/jbc.M001956200.
R. Kalluri, Cold Spring Harb. Symp. Quant. Biol., 67, 255 (2002); doi:10.1101/sqb.2002.67.255.
N. Floquet, S. Pasco, L. Ramont, P. Derreumaux, J.Y. Laronze, J.M. Nuzillard, F.X. Maquart, A.J. Alix and J.C. Monboisse, J. Biol. Chem., 279, 2091 (2004); doi:10.1074/jbc.M307736200.
Y. Maeshima, A. Sudhakar, J.C. Lively, K. Ueki, S. Kharbanda, C.R. Kahn, N. Sonenberg, R.O. Hynes and R. Kalluri, Science, 295, 140 (2002); doi:10.1126/science.1065298.
J. Thevenard, L. Ramont, J. Devy, B. Brassart, A. Dupont-Deshorgue, N. Floquet, L. Schneider, F. Ouchani, C. Terryn, F.X. Maquart, J.C. Monboisse and S. Brassart-Pasco, Int. J. Cancer., 126, 1055 (2010); doi:10.1002/ijc.24688.
Y. He, Y. Jiang, Y.-J. Li, X.-H. Liu, L. Zhang, L.-J. Liu, H. Shi, H.-N. Li, Y.-C. Ma and X.-M. Jin, J. Gastroenterol. Hepatol., 25, 935 (2010); doi:10.1111/j.1440-1746.2009.06209.x.
C.S. Boosani, A.K. Varma and A. Sudhakar, J. Cancer Sci. Ther., 2009, 8 (2010).