Inhibition of Plasmepsin V activity demonstrates its essential role in protein export, PfEMP1 display, and survival of malaria parasites.
Inhibition of Plasmepsin V activity demonstrates its essential role in protein export, PfEMP1 display, and survival of malaria parasites.
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DOI：
10.1371/journal.pbio.1001897
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发表时间：
2014-07
期刊：
影响因子：
9.8
通讯作者：
Boddey JA
中科院分区：
文献类型：
--
作者：
Sleebs BE;Lopaticki S;Marapana DS;O'Neill MT;Rajasekaran P;Gazdik M;Günther S;Whitehead LW;Lowes KN;Barfod L;Hviid L;Shaw PJ;Hodder AN;Smith BJ;Cowman AF;Boddey JA
疟疾蛋白酶Plasmepsin V的小分子抑制剂损害蛋白质输出和细胞重塑，减少寄生虫在人红细胞中的存活。疟疾寄生虫恶性疟原虫将数百种蛋白质输出到受感染的红细胞中，这些蛋白质参与细胞重塑和严重的毒性。输出机制涉及疟原虫输出元件（PEXEL），其是寄生虫蛋白酶Plasmepsin V（PMV）的切割位点。PMV基因是难删除的，这表明它是必不可少的，但缺乏明确的证据。在这里，我们产生了PEXEL模拟抑制剂，有效地阻断从恶性疟原虫和间日疟原虫分离的PMV的活性。恶性疟原虫中PMV活性的评估显示PEXEL切割协同发生，类似于信号肽酶。用抑制剂处理恶性疟原虫感染的红细胞引起PEXEL加工以及蛋白质输出的剂量依赖性抑制，包括红细胞表面上主要毒力粘附素PfEMP 1和细胞粘附的显示受损。抑制剂杀死寄生虫在滋养体阶段和敲减PMV增强敏感性的抑制剂，而过表达的PMV增加阻力。这提供了第一个直接的证据表明，PMV活性是必要的蛋白质输出疟原虫属。和寄生虫在人红细胞中的存活，并验证PMV作为抗疟疾药物靶标。为了在人类红细胞中生存，疟原虫必须输出一系列蛋白质，这些蛋白质重塑宿主细胞及其表面。这使得寄生虫能够从细胞外获得营养，并修改细胞表面以逃避宿主防御。蛋白质出口涉及蛋白水解裂解的疟原虫出口元件（PEXEL）的乙酰化蛋白酶Plasmepsin V.我们在这里报告的发展，密切模仿天然的PEXEL基板和阻断活性的疟原虫的恶性疟原虫和间日疟原虫的Plasmepsin V的小分子抑制剂。该抑制剂损害输出和细胞重塑，并在环状滋养体转变时杀死恶性疟原虫，提供了直接证据表明Plasmepsin V活性对于宿主内PEXEL蛋白的输出和寄生虫存活至关重要。这些发现证实Plasmepsin V是高度保守的抗疟药物靶标。
A small molecule inhibitor of the malarial protease Plasmepsin V impairs protein export and cellular remodeling, reducing parasite survival in human erythrocytes. The malaria parasite Plasmodium falciparum exports several hundred proteins into the infected erythrocyte that are involved in cellular remodeling and severe virulence. The export mechanism involves the Plasmodium export element (PEXEL), which is a cleavage site for the parasite protease, Plasmepsin V (PMV). The PMV gene is refractory to deletion, suggesting it is essential, but definitive proof is lacking. Here, we generated a PEXEL-mimetic inhibitor that potently blocks the activity of PMV isolated from P. falciparum and Plasmodium vivax. Assessment of PMV activity in P. falciparum revealed PEXEL cleavage occurs cotranslationaly, similar to signal peptidase. Treatment of P. falciparum–infected erythrocytes with the inhibitor caused dose-dependent inhibition of PEXEL processing as well as protein export, including impaired display of the major virulence adhesin, PfEMP1, on the erythrocyte surface, and cytoadherence. The inhibitor killed parasites at the trophozoite stage and knockdown of PMV enhanced sensitivity to the inhibitor, while overexpression of PMV increased resistance. This provides the first direct evidence that PMV activity is essential for protein export in Plasmodium spp. and for parasite survival in human erythrocytes and validates PMV as an antimalarial drug target. To survive within human red blood cells, malaria parasites must export a catalog of proteins that remodel the host cell and its surface. This enables parasites to acquire nutrients from outside the cell and to modify the cell surface in order to evade host defenses. Protein export involves proteolytic cleavage of the Plasmodium Export Element (PEXEL) by the aspartyl protease Plasmepsin V. We report here the development of a small molecule inhibitor that closely mimics the natural PEXEL substrate and blocks the activity of Plasmepsin V from the malarial parasites Plasmodium falciparum and Plasmodium vivax. The inhibitor impairs export and cellular remodeling and kills P. falciparum at the ring-trophozoite transition, providing direct evidence that Plasmepsin V activity is essential for export of PEXEL proteins and parasite survival within the host. These findings validate Plasmepsin V as a highly conserved antimalarial drug target.
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影响因子：
1.5
作者：
Deponte, Marcel;Hoppe, Heinrich C.;Przyborski, Jude M.
通讯作者：
Przyborski, Jude M.
DOI：
10.4269/ajtmh.2001.64.97
发表时间：
2001-01-01
影响因子：
3.3
作者：
Mendis, K;Sina, BJ;Carter, R
通讯作者：
Carter, R
DOI：
10.1111/tra.12049
发表时间：
2013-05
期刊：
Traffic (Copenhagen, Denmark)
影响因子：
--
作者：
Hsiao CH;Luisa Hiller N;Haldar K;Knoll LJ
通讯作者：
Knoll LJ
影响因子：
64.5
作者：
Maier AG;Rug M;O'Neill MT;Brown M;Chakravorty S;Szestak T;Chesson J;Wu Y;Hughes K;Coppel RL;Newbold C;Beeson JG;Craig A;Crabb BS;Cowman AF
通讯作者：
Cowman AF
影响因子：
120.1
作者：
Drag, Marcin;Salvesen, Guy S.
通讯作者：
Salvesen, Guy S.