Fernandes Frederico Francisco, Moraes Juliana Rodrigues, Dos Santos Jaqueline Leal, Soares Thiago Geraldo, Gouveia Vitor José Pinto, Matavel Alessandra C S, Borges William de Castro, Cordeiro Marta do Nascimento, Figueiredo Suely Gomes, Borges Márcia Helena
Laboratory of Proteomics and Arachnids, Research and Development Board, Ezequiel Dias Foundation, Belo Horizonte, MG, Brazil.
Laboratory of Enzymology and Proteomics, Department of Biological Sciences, Federal University of Ouro Preto, Ouro Preto, MG, Brazil.
J Venom Anim Toxins Incl Trop Dis. 2022 Feb 9;28:e20210042. doi: 10.1590/1678-9199-JVATITD-2021-0042. eCollection 2022.
Spider venoms induce different physio-pharmacological effects by binding with high affinity on molecular targets, therefore being of biotechnological interest. Some of these toxins, acting on different types of ion channels, have been identified in the venom of spiders of the genus , mainly from . In spite of the pharmaceutical potential demonstrated by toxins, there is limited information on molecules from venoms of the same genus, as their toxins remain poorly characterized. Understanding this diversity and clarifying the differences in the mechanisms of action of spider toxins is of great importance for establishing their true biotechnological potential. This prompted us to compare three different venoms of the genus: (Pn-V), (Pe-V) and (Pp-V).
Biochemical and functional comparison of the venoms were carried out by SDS-PAGE, HPLC, mass spectrometry, enzymatic activities and electrophysiological assays (whole-cell patch clamp).
The employed approach revealed that all three venoms had an overall similarity in their components, with only minor differences. The presence of a high number of similar proteins was evident, particularly toxins in the mass range of ~6.0 kDa. Hyaluronidase and proteolytic activities were detected in all venoms, in addition to isoforms of the toxins Tx1 and Tx2-6. All Tx1 isoforms blocked Nav1.6 ion currents, with slight differences.
Our findings showed that Pn-V, Pe-V and Pp-V are highly similar concerning protein composition and enzymatic activities, containing isoforms of the same toxins sharing high sequence homology, with minor modifications. However, these structural and functional variations are very important for venom diversity. In addition, our findings will contribute to the comprehension of the molecular diversity of the venoms of the other species from genus, exposing their biotechnological potential as a source for searching for new active molecules.
蜘蛛毒液通过与分子靶点高亲和力结合诱导不同的生理药理效应，因此具有生物技术研究价值。这些毒素中的一些作用于不同类型的离子通道，已在 属蜘蛛的毒液中被鉴定出来，主要来自 。尽管 毒素已显示出药物潜力，但关于同一属毒液中的分子信息有限，因为其毒素的特征仍不清楚。了解这种多样性并阐明蜘蛛毒素作用机制的差异对于确定它们真正的生物技术潜力至关重要。这促使我们比较 属的三种不同毒液： （Pn-V）、 （Pe-V）和 （Pp-V）。
通过SDS-PAGE、HPLC、质谱、酶活性和电生理测定（全细胞膜片钳）对毒液进行生化和功能比较。
所采用的方法表明，所有三种毒液的成分总体相似，只有微小差异。大量相似蛋白质的存在很明显，特别是在约6.0 kDa质量范围内的毒素。除了毒素Tx1和Tx2-6的同工型外，在所有毒液中都检测到了透明质酸酶和蛋白水解活性。所有Tx1同工型都阻断Nav1.6离子电流，只是略有差异。
我们的研究结果表明，Pn-V、Pe-V和Pp-V在蛋白质组成和酶活性方面高度相似，含有具有高度序列同源性的相同毒素的同工型，只是有微小修饰。然而，这些结构和功能变化对于毒液多样性非常重要。此外，我们的研究结果将有助于理解 属其他物种毒液的分子多样性，揭示它们作为寻找新活性分子来源的生物技术潜力。
