Division of Molecular Pathobiology, DCR-VPH, Vetsuisse Faculty, University of Bern, CH-3012 Bern, Switzerland.
Int J Parasitol. 2013 Feb;43(2):173-80. doi: 10.1016/j.ijpara.2012.10.017. Epub 2012 Nov 23.
The apicomplexan parasite, Theileria annulata, is the causative agent of tropical theileriosis, a devastating lymphoproliferative disease of cattle. The schizont stage transforms bovine leukocytes and provides an intriguing model to study host/pathogen interactions. The genome of T. annulata has been sequenced and transcriptomic data are rapidly accumulating. In contrast, little is known about the proteome of the schizont, the pathogenic, transforming life cycle stage of the parasite. Using one-dimensional (1-D) gel LC-MS/MS, a proteomic analysis of purified T. annulata schizonts was carried out. In whole parasite lysates, 645 proteins were identified. Proteins with transmembrane domains (TMDs) were under-represented and no proteins with more than four TMDs could be detected. To tackle this problem, Triton X-114 treatment was applied, which facilitates the extraction of membrane proteins, followed by 1-D gel LC-MS/MS. This resulted in the identification of an additional 153 proteins. Half of those had one or more TMD and 30 proteins with more than four TMDs were identified. This demonstrates that Triton X-114 treatment can provide a valuable additional tool for the identification of new membrane proteins in proteomic studies. With two exceptions, all proteins involved in glycolysis and the citric acid cycle were identified. For at least 29% of identified proteins, the corresponding transcripts were not present in the existing expressed sequence tag databases. The proteomics data were integrated into the publicly accessible database resource at EuPathDB (www.eupathdb.org) so that mass spectrometry-based protein expression evidence for T. annulata can be queried alongside transcriptional and other genomics data available for these parasites.
锥虫属寄生虫，环形泰勒虫，是热带泰勒虫病的病原体，这是一种毁灭性的牛淋巴增生性疾病。裂殖体阶段可转化牛白细胞，为研究宿主/病原体相互作用提供了一个有趣的模型。环形泰勒虫的基因组已经测序，转录组数据也在迅速积累。相比之下，裂殖体（寄生虫的致病、转化生命周期阶段）的蛋白质组知之甚少。通过一维（1-D）凝胶 LC-MS/MS，对纯化的环形泰勒虫裂殖体进行了蛋白质组分析。在整个寄生虫裂解物中，鉴定出 645 种蛋白质。具有跨膜结构域（TMD）的蛋白质代表性不足，并且无法检测到具有超过四个 TMD 的蛋白质。为了解决这个问题，应用了 Triton X-114 处理，这促进了膜蛋白的提取，然后进行 1-D 凝胶 LC-MS/MS。这导致鉴定出另外 153 种蛋白质。其中一半具有一个或多个 TMD，并且鉴定出 30 种具有超过四个 TMD 的蛋白质。这表明 Triton X-114 处理可以为蛋白质组学研究中鉴定新的膜蛋白提供有价值的附加工具。除了两个例外，所有参与糖酵解和柠檬酸循环的蛋白质都被鉴定出来。对于至少 29%的鉴定出的蛋白质，相应的转录本不存在于现有的表达序列标签数据库中。蛋白质组学数据已整合到公共可访问的数据库资源 EuPathDB（www.eupathdb.org）中，以便可以查询基于质谱的环形泰勒虫蛋白质表达证据，以及这些寄生虫可用的转录组和其他基因组学数据。
