Segura César, Cuesta-Astroz Yesid, Nunes-Batista Camila, Zalis Mariano, von Krüger Wanda Maria de Almeida, Mascarello Bisch Paulo
Grupo Malaria, Facultad de Medicina, Universidad de antioquia.
Laboratorio de Infectología y Parasitología Molecular, Hospital Clementino Fraga Filho, Universidad Federal de Rio de Janeiro.
Biomedica. 2014 Apr-Jun;34(2):237-49. doi: 10.1590/S0120-41572014000200010.
Despite efforts to control malaria, around 10% of the world population is at risk of acquiring this disease. Plasmodium falciparum accounts for the majority of severe cases and deaths. Malaria control programs have failed due to the therapeutic failure of first-line antimalarials and to parasite resistance. Thus, new and better therapeutic alternatives are required. Proteomic analysis allows determination of protein expression levels under drug pressure, leading to the identification of new therapeutic drug targets and their mechanisms of action.
The aim of this study was to analyze qualitatively the expression of P.falciparum trophozoite proteins (strain ITG2), after exposure to antimalarial drugs, through a proteomic approach.
In vitro cultured synchronized parasites were treated with quinine, mefloquine and the natural antiplasmodial diosgenone. Protein extracts were prepared and analyzed by two-dimensional electrophoresis. The differentially expressed proteins were selected and identified by MALDI-TOF mass spectrometry.
The following proteins were identified among those differentially expressed in the parasite in the presence of the drugs tested: enolase (PF10_0155), calcium-binding protein (PF11_0098), chaperonin (PFL0740c), the host cell invasion protein (PF10_0268) and proteins related to redox processes (MAL8P1.17). These findings are consistent with results of previous studies where the parasite was submitted to pressure with other antimalarial drugs.
The observed changes in the P. falciparum trophozoite protein profile induced by antimalarial drugs involved proteins mainly related to the general stress response.
尽管人们努力控制疟疾，但全球约10%的人口仍有感染这种疾病的风险。恶性疟原虫导致了大多数严重病例和死亡。由于一线抗疟药的治疗失败以及寄生虫耐药性，疟疾控制项目已告失败。因此，需要新的、更好的治疗选择。蛋白质组学分析能够确定药物压力下的蛋白质表达水平，从而识别新的治疗药物靶点及其作用机制。
本研究旨在通过蛋白质组学方法定性分析抗疟药物作用后恶性疟原虫滋养体蛋白（ITG2株）的表达。
用奎宁、甲氟喹和天然抗疟药薯蓣皂苷元处理体外培养的同步化寄生虫。制备蛋白质提取物并通过二维电泳进行分析。选择差异表达的蛋白质并通过基质辅助激光解吸电离飞行时间质谱进行鉴定。
在受试药物作用下，在寄生虫中差异表达的蛋白质中鉴定出以下蛋白质：烯醇化酶（PF10_0155）、钙结合蛋白（PF11_0098）、伴侣蛋白（PFL0740c）、宿主细胞入侵蛋白（PF10_0268）以及与氧化还原过程相关的蛋白质（MAL8P1.17）。这些发现与之前用其他抗疟药对该寄生虫施加压力的研究结果一致。
抗疟药物诱导的恶性疟原虫滋养体蛋白质谱变化涉及的蛋白质主要与一般应激反应相关。
