Vipera ammodytes (Va), is the European venomous snake of the greatest medical importance. We analyzed whole venom proteome of the subspecies V. ammodytes ammodytes (Vaa) from Serbia for the first time using the shotgun proteomics approach and identified 99 proteins belonging to four enzymatic families: serine protease (SVSPs), L-amino acid oxidase (LAAOs), metalloproteinases (SVMPs), group II phospholipase (PLA2s), and five nonenzymatic families: cysteine-rich secretory proteins (CRISPs), C-type lectins (snaclecs), growth factors -nerve (NGFs) and vascular endothelium (VEGFs), and Kunitz-type protease inhibitors (SPIs). Considerable enzymatic activity of LAAO, SVSPs, and SVMPs and a high acidic PLA2 activity was measured implying potential of Vaa to produce haemotoxic, myotoxic, neuro and cardiotoxic effects. Moreover, significant antimicrobial activity of Vaa venom against Gram-negative (Klebsiella pneumoniae, Pseudomonas aeruginosa) and Gram-positive bacteria (Staphylococcus aureus) was found. The crude venom shows considerable potential cytotoxic activity on the C6 and HL60 and a moderate level of potency on B16 cell lines. HeLa cells showed the same sensitivity, while DU 145 and PC-3 are less sensitive than as normal cell line. Our data demonstrated a high complexity of Vaa and considerable enzymatic, antibacterial and cytotoxic activity, implying a great medical potential of Vaa venom as a promising source for new antibacterial and cytostatic agents.

Vipera ammodytes ( Va )，是欧洲最重要的医学重要毒蛇。我们首次使用鸟枪蛋白质组学方法分析了来自塞尔维亚的亚种V. ammodytes ammodytes ( Vaa ) 的整个毒液蛋白质组，并鉴定了属于四个酶家族的 99 种蛋白质：丝氨酸蛋白酶 (SVSP)、L-氨基酸氧化酶 (LAAO) 、金属蛋白酶 (SVMP)、II 组磷脂酶 (PLA2) 和五个非酶家族：富含半胱氨酸的分泌蛋白 (CRISP)、C 型凝集素 (snaclecs)、生长因子 - 神经 (NGF) 和血管内皮 (VEGF)，以及Kunitz 型蛋白酶抑制剂 (SPI)。测量了 LAAO、SVSP 和 SVMP 的相当大的酶活性和高酸性 PLA2 活性，这意味着Vaa产生血液毒性、肌肉毒性、神经和心脏毒性作用。此外，发现Vaa毒液对革兰氏阴性菌（肺炎克雷伯菌、铜绿假单胞菌）和革兰氏阳性菌（金黄色葡萄球菌）具有显着的抗菌活性。粗毒液对 C6 和 HL60 显示出相当大的潜在细胞毒活性，对 B16 细胞系显示出中等水平的效力。HeLa 细胞表现出相同的敏感性，而 DU 145 和 PC-3 的敏感性低于正常细胞系。我们的数据显示了Vaa的高度复杂性以及相当大的酶、抗菌和细胞毒活性，这意味着Vaa具有巨大的医疗潜力 毒液作为新的抗菌剂和细胞抑制剂的有希望的来源。