Chang Long-Sen, Chen Ku-Chung, Lin Shinne-Ren, Huang Hsien-Bin
Institute of Biomedical Sciences, National Sun Yat-Sen Unversity-Kaohsiung Medical University Joint Center, National Sun Yat-Sen University, Kaohsiung 804, Taiwan, ROC.
Toxicon. 2006 Sep 15;48(4):429-36. doi: 10.1016/j.toxicon.2006.06.012. Epub 2006 Jun 29.
Three cytotoxin-like proteins from the venom of Ophiophagus hannah were isolated by a combination of ion exchange chromatography and reverse phase HPLC. Amino acid sequence analysis revealed that these proteins all consisted of 63 amino acids and shared approximate 50% and 56% sequence identity with Naja naja atra cardiotoxins and cardiotoxin-like basic proteins (CLBPs), respectively. CD spectra revealed that their secondary structure was dominated with beta-sheet as those noted with cardiotoxins and CLBPs. O. hannah cytotoxin-like protein exhibited a cell-lytic activity on SK-N-SH cells, but its activity was more weak than that noted for N. naja atra cardiotoxin 3. Alternatively, apoptotic cell death was induced by the addition of N. naja atra CLBP. Based on the sequence information with the toxin molecules, the functional residues and regions related to the differential activity with O. hannah cytotoxin-like protein, cardiotoxin and CLBP are discussed.
通过离子交换色谱和反相高效液相色谱相结合的方法，从眼镜王蛇毒液中分离出三种细胞毒素样蛋白。氨基酸序列分析表明，这些蛋白均由63个氨基酸组成，分别与眼镜蛇心脏毒素和心脏毒素样碱性蛋白（CLBPs）具有约50%和56%的序列同一性。圆二色光谱显示，它们的二级结构以β-折叠为主，与心脏毒素和CLBPs相同。眼镜王蛇细胞毒素样蛋白对SK-N-SH细胞具有细胞溶解活性，但其活性比眼镜蛇心脏毒素3弱。另外，添加眼镜蛇CLBP可诱导细胞凋亡。基于毒素分子的序列信息，讨论了与眼镜王蛇细胞毒素样蛋白、心脏毒素和CLBP活性差异相关的功能残基和区域。
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