Monogenea,水产养殖中普遍存在的寄生虫,对行业构成重大威胁,导致重大损失。目前的预防措施被证明是不够的,需要开发新的有效的抗寄生虫药物。在这次调查中,我们通过分析三代转录组数据获得了全长肌球蛋白cDNA序列,揭示了一个编码1938个氨基酸的5817个碱基的序列。随后,我们对肌球蛋白的二级和三级特征进行了建模和分析,精确定位运动域内的关键功能区域(氨基酸1-768)。该结构域的原核表达产生了87.44kDa的蛋白质,通过WesternBlotting确认为肌球蛋白。分子对接鉴定ASN439为参与牛肝素苷元与肌球蛋白结合的关键氨基酸残基,定点诱变证实的结果,确认这种相互作用的活跃腔。查尔酮和紫草素是从基于活性腔的天然药物分子库的虚拟筛中选择的。查尔酮和紫草素的EC50值分别为1.085mg/L和0.371mg/L,分别,肌球蛋白的相应IC50值为0.44mM和0.14mM。鉴于其优越的活性和结构,紫草素被选择用于进一步优化药物分子设计,最终发现了1,4-萘醌作为一种有效的抗寄生虫药。该化合物的EC50为0.047mg/L,LC50为0.23mg/L,TI指数为4.893。这些发现共同强调了紫草素和1,4-萘醌作为控制Gyrodactylus感染的替代化合物的潜力。药物化学的进一步优化有望开发更有效的1,4-萘醌类似物,通过组合或替代策略为未来的驱虫药控制提供了前景。
Monogenea, a prevalent parasite in aquaculture, poses significant threats to the industry, leading to substantial losses. Current preventive measures have proven insufficient, necessitating the development of novel and effective anti-parasitic drugs. In this investigation, we obtained the full-length
myosin cDNA sequence by analyzing three-generation transcriptome data, revealing a 5817-base sequence encoding 1938 amino acids. Subsequently, we modeled and analyzed the characteristics of the secondary and tertiary of
myosin, pinpointing the crucial functional region within the motor domain (amino acids 1-768). The prokaryotic expression of this domain yielded a protein of 87.44 kDa, confirmed as
myosin by Western Blotting. Molecular docking identified ASN439 as the key amino acid residue involved in arctigenin and
myosin binding, a result corroborated by site-directed mutagenesis, affirming the active cavity of this interaction. Chalcone and shikonin were chosen from a virtual sieve of molecular library of natural drugs based on the active cavity. Chalcone and shikonin exhibited EC50 values of 1.085 mg/L and 0.371 mg/L, respectively, with corresponding IC50 values for
myosin of 0.44 mM and 0.14 mM. Given its superior activity and structure, shikonin was selected for further optimization of drug molecule design, culminating in the discovery of 1,4-naphthoquinone as a potent antiparasitic agent. This compound demonstrated an EC50 of 0.047 mg/L, LC50 of 0.23 mg/L, and a TI index of 4.893. These findings collectively highlight the potential of shikonin and 1,4-naphthoquinone as alternative compounds to control Gyrodactylus infections. Further optimization of medicinal chemistry holds promise for the development of more potent 1,4-naphthoquinone analogues, offering prospects for future anthelmintic control through combinatorial or replacement strategies.