The Prenylated Proteome of Plasmodium falciparum Reveals Pathogen-specific Prenylation Activity and Drug Mechanism-of-action
The Prenylated Proteome of Plasmodium falciparum Reveals Pathogen-specific Prenylation Activity and Drug Mechanism-of-action
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DOI：
10.1074/mcp.m116.064550
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发表时间：
2017-04-01
影响因子：
7
通讯作者：
Yeh, Ellen
中科院分区：
文献类型：
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作者：
Gisselberg, Jolyn E.;Zhang, Lichao;Yeh, Ellen
疟原虫含有几个独特的膜隔室，其中异戊二烯化蛋白可能在发病机制中发挥重要作用。蛋白质异戊烯化也被提议作为抗疟疾药物靶点，因为法尼基转移酶抑制剂引起血液阶段疟原虫的有效生长抑制。然而，介导抗疟活性的特定异戊二烯化蛋白质尚未被鉴定。鉴于新寄生虫生物学的潜力和阐明药物作用机制的潜力，我们使用炔标记的异戊烯类似物来特异性富集寄生虫异戊烯化蛋白质，对血液期恶性疟原虫中的异戊烯化蛋白质组进行了大规模鉴定。确定了20个高置信度的候选者，包括病原体特异性异戊烯化活性的几个例子。一种独特的寄生虫异戊二烯化蛋白是含有FYVE的卷曲螺旋蛋白（FCP），其仅在疟原虫和相关顶复门寄生虫中保守，并定位于寄生虫食物泡。FCP靶向该寄生虫特异性隔室依赖于其CaaX基序的异戊烯化，因为异戊烯化位点的突变引起细胞溶质错误定位。我们还表明，PfRab 5 b，其中缺乏C-末端半胱氨酸，这是唯一已知的网站的Rab GTdR修饰，是异戊二烯化。最后，我们表明，THQ类的法呢基转移酶抑制剂废除FCP异戊二烯化，并导致其错误定位，提供了第一个演示的抗疟疾法呢基转移酶抑制剂破坏一个特定的异戊二烯化蛋白。总而言之，这些研究结果确定了异戊二烯化蛋白，揭示了独特的寄生虫生物学，并用于评价异戊二烯基转移酶抑制剂的抗疟药物开发。
Plasmodium parasites contain several unique membrane compartments in which prenylated proteins may play important roles in pathogenesis. Protein prenylation has also been proposed as an antimalarial drug target because farnesyltransferase inhibitors cause potent growth inhibition of blood-stage Plasmodium. However, the specific prenylated proteins that mediate antimalarial activity have yet to be identified. Given the potential for new parasite biology and elucidating drug mechanism-of-action, we performed a large-scale identification of the prenylated proteome in blood-stage P. falciparum parasites using an alkyne-labeled prenyl analog to specifically enrich parasite prenylated proteins. Twenty high-confidence candidates were identified, including several examples of pathogen-specific prenylation activity. One unique parasite prenylated protein was FYVE-containing coiled-coil protein (FCP), which is only conserved in Plasmodium and related Apicomplexan parasites and localizes to the parasite food vacuole. Targeting of FCP to this parasite-specific compartment was dependent on prenylation of its CaaX motif, as mutation of the prenylation site caused cytosolic mislocalization. We also showed that PfRab5b, which lacks C-terminal cysteines that are the only known site of Rab GTPase modification, is prenylated. Finally, we show that the THQ class of farnesyltransferase inhibitors abolishes FCP prenylation and causes its mislocalization, providing the first demonstration of a specific prenylated protein disrupted by antimalarial farnesyl transferase inhibitors. Altogether, these findings identify prenylated proteins that reveal unique parasite biology and are useful for evaluating prenyltransferase inhibitors for antimalarial drug development.