Conlon J Michael, Prajeep Manju, Mechkarska Milena, Arafat Kholoud, Attoub Samir, Adem Abdu, Pla Davinia, Calvete Juan J
Department of Biochemistry, College of Medicine and Health Sciences, United Arab Emirates University, Al Ain, United Arab Emirates.
Department of Pharmacology, College of Medicine and Health Sciences, United Arab Emirates University, Al Ain, United Arab Emirates.
J Venom Res. 2014 Jun 19;5:16-21. eCollection 2014.
Two structurally related (48.6% amino acid sequence identity) peptides with cytotoxic activity against human non-small cell lung adenocarcinoma A549 cells were purified from the venom of the Eastern green mamba Dendroaspis angusticeps using reversed phase HPLC. The peptides were identified as members of the three-finger superfamily of snake toxins by mass fingerprinting of tryptic digests. The more potent peptide (LC50 against A549 cells = 56±4µg/ml) was identical to the previously described toxin C13S1C1 and the less active peptide (LC50 against A549 cells = 106±5µg/ml) was identical to toxin F-VIII. Toxin C13S1C1 was also cytotoxic against breast adenocarcinoma MDA-MB-231 cells (LC50 = 62±2µg/ml) and colorectal adenocarcinoma HT-29 cells (LC50 = 110±4µg/ml). Although the peptide was appreciably less hemolytic activity against human erythrocytes (LC50 >600µg/ml), it was cytotoxic to human umbilical vein endothelial HUVEC cells (57±3µg/ml) indicating no differential activity against cell lines derived from neoplastic tissues. Toxin F-VIII was not cytotoxic to MDA-MB-231, HT-29 cells, and HUVEC cells at concentrations up to 300µg/ml and was not hemolytic at concentrations up to 1mg/ml. Neither peptide inhibited growth of reference strains of Escherichia coli or Staphylococcus aureus (MIC values >200μg/ml).
利用反相高效液相色谱法从东部绿曼巴蛇(Dendroaspis angusticeps)的毒液中纯化出两种对人非小细胞肺腺癌A549细胞具有细胞毒性活性的结构相关肽(氨基酸序列同一性为48.6%)。通过胰蛋白酶消化产物的质谱指纹图谱鉴定这些肽为蛇毒素三指超家族的成员。活性更强的肽(对A549细胞的半数致死浓度=56±4μg/ml)与先前描述的毒素C13S1C1相同,活性较弱的肽(对A549细胞的半数致死浓度=106±5μg/ml)与毒素F-VIII相同。毒素C13S1C1对乳腺腺癌MDA-MB-231细胞(半数致死浓度=62±2μg/ml)和结肠腺癌HT-29细胞(半数致死浓度=110±4μg/ml)也具有细胞毒性。尽管该肽对人红细胞的溶血活性明显较低(半数致死浓度>600μg/ml),但它对人脐静脉内皮HUVEC细胞具有细胞毒性(57±3μg/ml),表明对源自肿瘤组织的细胞系没有差异活性。毒素F-VIII在浓度高达300μg/ml时对MDA-MB-231、HT-29细胞和HUVEC细胞没有细胞毒性,在浓度高达1mg/ml时没有溶血活性。这两种肽均未抑制大肠杆菌或金黄色葡萄球菌参考菌株的生长(最低抑菌浓度值>200μg/ml)。