Blocking effect of a biotinylated protease inhibitor on the egress of Plasmodium falciparum merozoites from infected red blood cells
Blocking effect of a biotinylated protease inhibitor on the egress of Plasmodium falciparum merozoites from infected red blood cells
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DOI：
10.1515/bc.2005.059
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发表时间：
2005-05-01
影响因子：
3.7
通讯作者：
Leippe, M
中科院分区：
文献类型：
--
作者：
Gelhaus, C;Vicik, R;Leippe, M
疟疾寄生虫恶性疟原虫侵入人体红细胞。在感染新的红细胞之前，裂殖子必须离开宿主细胞进入血浆。关于排出机制的知识很少，但据认为蛋白酶基本上参与了这一步骤。我们引入了生物素化的氮丙啶-2，3-二羧酸二苄酯（bADA）作为不可逆的半胱氨酸蛋白酶抑制剂，以研究裂殖子释放的机制，并确定参与的蛋白酶。该化合物作用于消化泡和宿主细胞胞质溶胶中的寄生虫蛋白，如通过荧光显微镜所判断的。抑制剂阻断宿主细胞膜的破裂，导致成簇的裂殖子结构，如通过免疫电子显微镜所证明的。有趣的是，bADA没有防止破裂的寄生虫空泡膜（PVM），围绕寄生虫在红细胞内成熟的时期。该化合物似乎是一个有价值的模板，用于开发针对单个疟原虫蛋白酶的抑制剂，这将是有用的工具来剖析裂殖子释放过程的分子机制，从而开发有效的抗疟疾药物。
The malaria parasite Plasmodium falciparum invades human red blood cells. Before infecting new erythrocytes, the merozoites have to exit their host cell to get into the blood plasma. Knowledge about the mechanism of egress is scarce, but it is thought that proteases are basically involved in this step. We have introduced a biotinylated dibenzyl aziridine-2,3-dicarboxylate (bADA) as an irreversible cysteine protease inhibitor to study the mechanism of merozoite release and to identify the proteases involved. The compound acts on parasite proteins in the digestive vacuole and in the host cell cytosol, as judged by fluorescence microscopy. The inhibitor blocks rupture of the host cell membrane, leading to clustered merozoite structures, as evidenced by immunoelectron microscopy. Interestingly, bADA did not prevent rupture of the parasitophorous vacuole membrane (PVM) that surrounds the parasite during the period of intraerythrocytic maturation. The compound appears to be a valuable template for the development of inhibitors specific for individual plasmodial proteases, which would be useful tools to dissect the molecular mechanisms underlying the process of merozoite release and consequently to develop potent antimalarial drugs.