Osipov Alexey V, Starkov Vladislav G, Tsetlin Victor I, Utkin Yuri N
Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy of Sciences, Moscow 117997, Russia.
Int J Mol Sci. 2025 May 1;26(9):4291. doi: 10.3390/ijms26094291.
Three-finger toxins (TFTs), including neurotoxins and cytotoxins, form one of the largest families of snake venom proteins and interact with various biological targets. Neurotoxins target proteinaceous receptors while cytotoxins interact mainly with the lipids of cell membranes and to a lesser extent with carbohydrates. However, no data about the interaction of TFTs with nucleic acids can be found. To detect this interaction, we applied spectrophotometry, ion-paired HPLC and electrophoretic mobility shift assay (EMSA). Using spectrophotometry, we found that TFTs from cobra venom increased the optical density of an RNA solution in a time-dependent manner indicating toxin interaction with RNA. A decrease in the net negative charge of the RNA molecule upon interaction with neurotoxin II from cobra venom was revealed by ion-pair HPLC. EMSA showed decreased electrophoretic mobility of both RNA and DNA upon addition of different TFTs including the non-conventional cobra toxin WTX and water-soluble recombinant human three-finger protein lynx1. We suggest that the interaction with nucleic acids may be a common property of TFTs, and some biological effects of TFTs, for example, cytotoxin-induced apoptosis in cancer cell lines, may be mediated by interaction with nucleic acids.
三指毒素（TFTs），包括神经毒素和细胞毒素，构成蛇毒蛋白中最大的家族之一，并与多种生物靶点相互作用。神经毒素作用于蛋白质受体，而细胞毒素主要与细胞膜脂质相互作用，与碳水化合物的相互作用较弱。然而，尚未发现有关TFTs与核酸相互作用的数据。为了检测这种相互作用，我们应用了分光光度法、离子对高效液相色谱法和电泳迁移率变动分析（EMSA）。通过分光光度法，我们发现眼镜蛇毒中的TFTs以时间依赖性方式增加了RNA溶液的光密度，表明毒素与RNA发生了相互作用。离子对高效液相色谱法显示，与眼镜蛇毒中的神经毒素II相互作用后，RNA分子的净负电荷减少。EMSA表明，添加不同的TFTs（包括非传统眼镜蛇毒素WTX和水溶性重组人三指蛋白lynx1）后，RNA和DNA的电泳迁移率均降低。我们认为，与核酸的相互作用可能是TFTs的共同特性，并且TFTs的一些生物学效应，例如细胞毒素诱导癌细胞系凋亡，可能是由与核酸的相互作用介导的。
