Institute of Parasitology, Macdonald Campus, McGill University , 21111 Lakeshore Road, Saine-Anne-de-Bellevue, Québec H9X 3V9, Canada.
University of Victoria -Genome British Columbia Proteomics Centre , #3101-4464 Markham Street, Vancouver Island Technology Park, Victoria, British Columbia V8Z7X8, Canada.
J Proteome Res. 2018 Mar 2;17(3):1194-1215. doi: 10.1021/acs.jproteome.7b00817. Epub 2018 Feb 5.
To facilitate a greater understanding of the biological processes in the medically important Leishmania donovani parasite, a combination of differential and density-gradient ultracentrifugation techniques were used to achieve a comprehensive subcellular fractionation of the promastigote stage. An in-depth label-free proteomic LC-MS/MS analysis of the density gradients resulted in the identification of ∼50% of the Leishmania proteome (3883 proteins detected), which included ∼645 integral membrane proteins and 1737 uncharacterized proteins. Clustering and subcellular localization of proteins was based on a subset of training Leishmania proteins with known subcellular localizations that had been determined using biochemical, confocal microscopy, or immunoelectron microscopy approaches. This subcellular map will be a valuable resource that will help dissect the cell biology and metabolic processes associated with specific organelles of Leishmania and related kinetoplastids.
为了更深入地了解医学上重要的利什曼原虫中的生物学过程，采用差速和密度梯度超速离心技术相结合的方法，对前鞭毛体阶段进行了全面的亚细胞分级分离。对密度梯度的无标记蛋白质组学 LC-MS/MS 分析深入研究，鉴定出约 50%的利什曼原虫蛋白质组（检测到 3883 种蛋白质），其中包括约 645 种完整的膜蛋白和 1737 种未鉴定的蛋白质。蛋白质的聚类和亚细胞定位是基于一组具有已知亚细胞定位的训练利什曼原虫蛋白质，这些蛋白质的亚细胞定位是使用生化、共聚焦显微镜或免疫电子显微镜方法确定的。这个亚细胞图谱将是一个有价值的资源，有助于剖析与利什曼原虫和相关动基体门生物特定细胞器相关的细胞生物学和代谢过程。
