Gold nanoparticles (GNPs) are one of the most widely used nanomaterials in various fields. Especially, the unique chemical and physical properties make them as the promising candidates in drug target identification, unfortunately, little is known about their application in parasites. In this paper, GNPs were employed as new solid support to identify drug targets of natural bioactive compound arctigenin (ARG) against fish monogenean parasite Gyrodactylus kobayashi. Before target identification, GNPs with ARG on the surface showed the ability to enter the live parasites even the nucleus or mitochondria, which made the bound compounds capable of contacting directly with target proteins located anywhere of the parasites. At the same time, chemically modified compound remained the anthelminthic efficacy against G. kobayashii. The above results both provide assurance on the reliability of using GNPs for drug target-binding specificity. Subsequently, by interrogating the cellular proteome in parasite lysate, myosin-2 and UNC-89 were identified as the potential direct target proteins of ARG in G. kobayashii. Moreover, results of RNA-seq transcriptomics and iTRAQ proteomics indicated that myosin-2 expressions were down-regulated after ARG bath treatment both in transcript and protein levels, but for UNC-89, only in mRNA level. Myosin-2 is an important structural muscle protein expressed in helminth tegument and its identification as our target will enable further inhibitor optimization towards future drug discovery. Furthermore, our findings demonstrate the power of GNPs to be readily applied to other parasite drugs of unknown targets, facilitating more broadly therapeutic drug design in any pathogen or disease model.

中文翻译：

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使用金纳米探针对蛋白质组进行询问，以鉴定鱼类寄生虫中反萃素的靶标

金纳米颗粒（GNP）是各个领域中使用最广泛的纳米材料之一。特别是，独特的化学和物理性质使其成为药物靶标鉴定中很有希望的候选者，不幸的是，对其在寄生虫中的应用知之甚少。在本文中，GNP被用作新的固体支持物，以鉴定天然生物活性复合物Arctigenin（ARG）对抗鱼类单基因寄生虫Gyrodactylus kobayashi的药物靶标。在鉴定靶标之前，表面带有ARG的GNPs能够进入活的寄生虫，甚至进入细胞核或线粒体，这使结合的化合物能够直接与位于寄生虫任何地方的靶蛋白接触。同时，化学修饰的化合物仍然具有驱除小林葛根的驱虫药的功效。以上结果均为使用GNPs用于药物靶标结合特异性的可靠性提供了保证。随后，通过询问寄生虫裂解物中的细胞蛋白质组，肌球蛋白2和UNC-89被鉴定为小林志贺菌中ARG的潜在直接靶蛋白。此外，RNA-seq转录组学和iTRAQ蛋白质组学的结果表明，肌球蛋白2表达在ARG浴处理后在转录和蛋白水平上均被下调，而对于UNC-89，仅在mRNA水平上被下调。肌球蛋白2是在蠕虫皮中表达的一种重要的结构性肌肉蛋白，将其鉴定为我们的靶标将使进一步的抑制剂优化成为未来药物开发的目标。此外，我们的发现表明，GNP可以很容易地应用于未知目标的其他寄生虫药物，