Liverpool School of Tropical Medicine, Pembroke Place, Liverpool, L3 5QA, UK.
The Francis Crick Institute, 1 Midland Road, London, NW1 1AT, UK.
Malar J. 2018 Aug 13;17(1):293. doi: 10.1186/s12936-018-2445-8.
To understand more about changes to the molecular components that occur when host endothelium interacts with Plasmodium falciparum-infected erythrocytes, a combined technique of protein separation (1D Blue-Native electrophoresis) and mass spectrometry of infected erythrocytes with endothelial cells (EC) in a co-culture system has been used.
Native proteins were extracted from co-cultures and identified by mass spectrometry. Proteomic data from different parasite strains, either adhesion proficient (to endothelial cells) or non-adherent, were analysed in parallel to reveal protein associations linked to cytoadherence. Informatic approaches were developed to facilitate this comparison.
Blue-Native gel separation and LC/MS/MS identification revealed major differences in samples produced from endothelial cell co-culture with adherent and non-adherent parasite strains. This approach enabled us to identify protein associations seen only with the adhesion proficient parasite strain.
The combination of proteomic and analytical approaches has identified differences between adherent and non-adherent parasite lines in co-culture with EC, providing potential candidates for complexes or associations formed during cytoadherence involved in cell structure, signalling and apoptosis.
为了更深入地了解宿主内皮细胞与感染疟原虫的红细胞相互作用时发生的分子成分变化，我们采用了一种联合技术，即蛋白分离（1D 蓝色天然电泳）和共培养系统中感染红细胞与内皮细胞（EC）的质谱分析。
从共培养物中提取天然蛋白，并通过质谱进行鉴定。对具有不同黏附能力（与内皮细胞黏附）或不黏附能力的寄生虫株的蛋白质组学数据进行平行分析，以揭示与细胞黏附相关的蛋白质关联。开发了信息学方法来促进这种比较。
蓝色天然凝胶分离和 LC/MS/MS 鉴定显示，来自与黏附性寄生虫株和非黏附性寄生虫株共培养的内皮细胞的样本之间存在明显差异。这种方法使我们能够鉴定仅与黏附性寄生虫株相关的蛋白质关联。
蛋白质组学和分析方法的结合，鉴定了共培养中黏附性和非黏附性寄生虫系之间的差异，为细胞结构、信号转导和细胞凋亡等细胞黏附过程中形成的复合物或关联提供了潜在的候选物。
