Zhou Chun-Xue, Zhu Xing-Quan, Elsheikha Hany M, He Shuai, Li Qian, Zhou Dong-Hui, Suo Xun
National Animal Protozoa Laboratory and College of Veterinary Medicine, China Agricultural University, Beijing 100193, PR China; State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province 730046, PR China.
State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Veterinary Parasitology of Gansu Province, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou, Gansu Province 730046, PR China.
J Proteomics. 2016 Oct 4;148:12-9. doi: 10.1016/j.jprot.2016.07.010. Epub 2016 Jul 12.
Toxoplasma gondii is a medically and economically important protozoan parasite. However, the molecular mechanisms of its sporulation remain largely unknown. Here, we applied iTRAQ coupled with 2D LC-MS/MS proteomic analysis to investigate the proteomic expression profile of T. gondii oocysts during sporulation. Of the 2095 non-redundant proteins identified, 587 were identified as differentially expressed proteins (DEPs). Based on Gene Ontology enrichment and KEGG pathway analyses the majority of these DEPs were found related to the metabolism of amino acids, carbon and energy. Protein interaction network analysis generated by STRING identified ATP-citrate lyase (ACL), GMP synthase, IMP dehydrogenase (IMPDH), poly (ADP-ribose) glycohydrolase (PARG), and bifunctional dihydrofolate reductase-thymidylate synthase (DHFR-TS) as the top five hubs. We also identified 25 parasite virulence factors that were expressed at relatively high levels in sporulated oocysts compared to non-sporulated oocysts, which might contribute to the infectivity of mature oocysts. Considering the importance of oocysts in the dissemination of toxoplasmosis these findings may help in the search of protein targets with a key role in infectiousness and ecological success of oocysts, creating new opportunities for the development of better means for disease prevention.
The development of new preventative interventions against T. gondii infection relies on an improved understanding of the proteome and chemical pathways of this parasite. To identify proteins required for the development of environmentally resistant and infective T. gondii oocysts, we compared the proteome of non-sporulated (immature) oocysts with the proteome of sporulated (mature, infective) oocysts. iTRAQ 2D-LC-MS/MS analysis revealed proteomic changes that distinguish non-sporulated from sporulated oocysts. Many of the differentially expressed proteins were involved in metabolic pathways and 25 virulence factors were identified upregulated in the sporulated oocysts. This work provides the first quantitative characterization of the proteomic variations that occur in T. gondii oocyst stage during sporulation.
刚地弓形虫是一种在医学和经济上具有重要意义的原生动物寄生虫。然而，其孢子形成的分子机制在很大程度上仍不清楚。在此，我们应用iTRAQ结合二维液相色谱-串联质谱蛋白质组学分析来研究刚地弓形虫卵囊在孢子形成过程中的蛋白质组表达谱。在鉴定出的2095种非冗余蛋白质中，有587种被鉴定为差异表达蛋白（DEP）。基于基因本体富集和KEGG通路分析，发现这些DEP中的大多数与氨基酸、碳和能量代谢有关。通过STRING生成的蛋白质相互作用网络分析确定，ATP-柠檬酸裂解酶（ACL）、鸟苷酸合酶、肌苷酸脱氢酶（IMPDH）、聚（ADP-核糖）糖苷水解酶（PARG）和双功能二氢叶酸还原酶-胸苷酸合酶（DHFR-TS）为前五个枢纽蛋白。我们还鉴定出25种寄生虫毒力因子，与未孢子化卵囊相比，它们在孢子化卵囊中表达水平相对较高，这可能有助于成熟卵囊的感染性。考虑到卵囊在弓形虫病传播中的重要性，这些发现可能有助于寻找在卵囊的感染性和生态成功中起关键作用的蛋白质靶点，为开发更好的疾病预防手段创造新机会。
针对刚地弓形虫感染开发新的预防干预措施依赖于对该寄生虫蛋白质组和化学途径的深入了解。为了鉴定形成具有环境抗性和感染性的刚地弓形虫卵囊所需的蛋白质，我们比较了未孢子化（不成熟）卵囊和孢子化（成熟、感染性）卵囊的蛋白质组。iTRAQ二维液相色谱-串联质谱分析揭示了区分未孢子化卵囊和孢子化卵囊的蛋白质组变化。许多差异表达蛋白参与代谢途径，并且在孢子化卵囊中鉴定出25种上调的毒力因子。这项工作首次对刚地弓形虫卵囊阶段在孢子形成过程中发生的蛋白质组变化进行了定量表征。
