School of Health and Biomedical Sciences, RMIT University, Bundoora, VIC, Australia.
Department of Veterinary Biosciences, Melbourne Veterinary School, The University of Melbourne, Parkville, VIC, Australia.
Parasit Vectors. 2022 Jul 30;15(1):273. doi: 10.1186/s13071-022-05400-w.
Gaining insight into molecular signalling pathways of socioeconomically important parasitic nematodes has implications for understanding their molecular biology and for developing novel anthelmintic interventions.
Here, we evaluated the use of a human antibody-based microarray to explore conserved elements of the signalome in the barber's pole worm Haemonchus contortus. To do this, we prepared extracts from mixed-sex (female and male) adult worms and third-stage larvae (L3s), incubated these extracts on the antibody microarray and then measured the amounts of antibody-bound proteins ('signal intensity').
In total, 878 signals were classified into two distinct categories: signals that were higher for adults than for larvae of H. contortus (n = 376), and signals that were higher for larvae than for adults of this species (n = 502). Following a data-filtering step, high confidence ('specific') signals were obtained for subsequent analyses. In total, 39 pan-specific signals (linked to antibodies that recognise target proteins irrespective of their phosphorylation status) and 65 phosphorylation-specific signals were higher in the adult stage, and 82 pan-specific signals and 183 phosphorylation-specific signals were higher in L3s. Thus, notably more signals were higher in L3s than in the adult worms. Using publicly available information, we then inferred H. contortus proteins that were detected (with high confidence) by specific antibodies directed against human homologues, and revealed relatively high structural conservation between the two species, with some variability for select proteins. We also in silico-matched 763 compound structures (listed in the DrugBank and Kinase SARfari public databases) to four H. contortus proteins (designated HCON_00005760, HCON_00079680, HCON_00013590 and HCON_00105100).
We conclude that the present antibody-based microarray provides a useful tool for comparative analyses of signalling pathways between/among developmental stages and/or species, as well as opportunities to explore nematocidal target candidates in H. contortus and related parasites.
深入了解具有社会经济重要性的寄生线虫的分子信号通路，对于理解它们的分子生物学和开发新型驱虫干预措施具有重要意义。
在这里，我们评估了使用基于人抗体的微阵列来探索旋毛虫 Haemonchus contortus 信号组中保守元件的方法。为此，我们从混合性别（雌性和雄性）成虫和第三期幼虫（L3）中制备提取物，将这些提取物孵育在抗体微阵列上，然后测量抗体结合蛋白的量（“信号强度”）。
总共，878 个信号被分为两类：一类是成虫的信号高于旋毛虫的幼虫（n=376），另一类是幼虫的信号高于该物种的成虫（n=502）。经过数据过滤步骤，获得了后续分析的高可信度（“特异性”）信号。总共，39 个泛特异性信号（与识别目标蛋白的抗体相关，无论其磷酸化状态如何）和 65 个磷酸化特异性信号在成虫阶段较高，82 个泛特异性信号和 183 个磷酸化特异性信号在 L3 阶段较高。因此，幼虫阶段的信号明显高于成虫阶段。然后，我们使用公开可用的信息推断出被特定针对人类同源物的抗体检测到（具有高可信度）的旋毛虫蛋白，并揭示了这两个物种之间相对较高的结构保守性，对于某些蛋白质则存在一些可变性。我们还在计算机上匹配了 763 种化合物结构（列在 DrugBank 和 Kinase SARfari 公共数据库中）到四个旋毛虫蛋白（指定为 HCON_00005760、HCON_00079680、HCON_00013590 和 HCON_00105100）。
我们得出结论，本基于抗体的微阵列为比较分析发育阶段之间/之间以及物种之间的信号通路提供了有用的工具，以及探索旋毛虫和相关寄生虫中的驱虫靶标候选物的机会。
