Yan Shuai, Tang Xiaochao, Yu Dianmei, Wang Haiyan, Meng Wenwen, Tang Pingping, Wang Xianchun
Key Laboratory of Protein Chemistry and Developmental Biology of Ministry of Education, College of Life Sciences, Hunan Normal University, Changsha 410081, Hunan, China.
Sheng Wu Gong Cheng Xue Bao. 2021 Feb 25;37(2):635-645. doi: 10.13345/j.cjb.200245.
One of the distinct characters of Latrodectus tredecimguttatus is that its toxic components exist not only in the venomous glands, but also in the tissues outside the venomous glands and even in the eggs. Investigation on the toxins outside the venomous glands can deepen our understanding of spider toxins and discover new lead molecules with important application prospects. In order to explore the low-abundance proteinaceous toxins in the L. tredecimguttatus eggs, we used bioinformatic strategies to mine a gene sequence encoding a peptide toxin from the transcriptome of L. tredecimguttatus eggs, and then heterologously expressed the gene successfully with a 3'-RACE combined with nest PCR strategy. Biological activity analyses indicated that the expressed peptide toxin, named latroeggtoxin-Ⅵ (LETX-Ⅵ), could inhibit Na⁺ channel currents in ND7/23 cells and promote dopamine release from PC12 cells, without obvious toxicity against Periplaneta americana and bacteria as well as fungi including Staphylococcus aureus and Candida albicans, demonstrating that LETX-Ⅵ is a mammal-specific neurotoxin with a potential application prospect in development of the tool reagents for neurobiological study and the drugs for treating related diseases.
红斑寇蛛的一个显著特征是其毒性成分不仅存在于毒腺中，还存在于毒腺外的组织甚至卵中。对毒腺外毒素的研究可以加深我们对蜘蛛毒素的理解，并发现具有重要应用前景的新先导分子。为了探索红斑寇蛛卵中的低丰度蛋白质毒素，我们利用生物信息学策略从红斑寇蛛卵的转录组中挖掘出一个编码肽毒素的基因序列，然后通过3'-RACE结合巢式PCR策略成功地对该基因进行了异源表达。生物活性分析表明，表达的肽毒素命名为latroeggtoxin-Ⅵ（LETX-Ⅵ），可以抑制ND7/23细胞中的Na⁺通道电流，并促进PC12细胞释放多巴胺，对美洲大蠊、细菌以及包括金黄色葡萄球菌和白色念珠菌在内的真菌没有明显毒性，表明LETX-Ⅵ是一种哺乳动物特异性神经毒素，在神经生物学研究工具试剂和相关疾病治疗药物的开发中具有潜在的应用前景。
