Tham Wai-Hong, Lim Nicholas T Y, Weiss Greta E, Lopaticki Sash, Ansell Brendan R E, Bird Megan, Lucet Isabelle, Dorin-Semblat Dominique, Doerig Christian, Gilson Paul R, Crabb Brendan S, Cowman Alan F
The Walter & Eliza Hall Institute of Medical Research, Parkville, Victoria, Australia.
Department of Medical Biology, The University of Melbourne, Parkville, Victoria, Australia.
PLoS Pathog. 2015 Dec 22;11(12):e1005343. doi: 10.1371/journal.ppat.1005343. eCollection 2015 Dec.
The most severe form of malaria in humans is caused by the protozoan parasite Plasmodium falciparum. The invasive form of malaria parasites is termed a merozoite and it employs an array of parasite proteins that bind to the host cell to mediate invasion. In Plasmodium falciparum, the erythrocyte binding-like (EBL) and reticulocyte binding-like (Rh) protein families are responsible for binding to specific erythrocyte receptors for invasion and mediating signalling events that initiate active entry of the malaria parasite. Here we have addressed the role of the cytoplasmic tails of these proteins in activating merozoite invasion after receptor engagement. We show that the cytoplasmic domains of these type 1 membrane proteins are phosphorylated in vitro. Depletion of PfCK2, a kinase implicated to phosphorylate these cytoplasmic tails, blocks P. falciparum invasion of red blood cells. We identify the crucial residues within the PfRh4 cytoplasmic domain that are required for successful parasite invasion. Live cell imaging of merozoites from these transgenic mutants show they attach but do not penetrate erythrocytes implying the PfRh4 cytoplasmic tail conveys signals important for the successful completion of the invasion process.
人类疟疾最严重的形式是由原生动物寄生虫恶性疟原虫引起的。疟原虫的侵入形式被称为裂殖子,它利用一系列与宿主细胞结合的寄生虫蛋白来介导侵入。在恶性疟原虫中,红细胞结合样(EBL)和网织红细胞结合样(Rh)蛋白家族负责与特定的红细胞受体结合以进行侵入,并介导引发疟原虫主动进入的信号事件。在这里,我们探讨了这些蛋白的细胞质尾巴在受体结合后激活裂殖子侵入中的作用。我们表明,这些1型膜蛋白的细胞质结构域在体外被磷酸化。PfCK2是一种与这些细胞质尾巴磷酸化有关的激酶,其缺失会阻断恶性疟原虫对红细胞的侵入。我们确定了PfRh4细胞质结构域中成功进行寄生虫侵入所需的关键残基。对这些转基因突变体的裂殖子进行活细胞成像显示,它们附着但不穿透红细胞,这意味着PfRh4细胞质尾巴传递了对成功完成侵入过程很重要的信号。