Protein sorting in Plasmodium falciparum-infected red blood cells permeabilized with the pore-forming protein streptolysin O
Protein sorting in Plasmodium falciparum-infected red blood cells permeabilized with the pore-forming protein streptolysin O
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DOI：
10.1042/bj3150307
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发表时间：
1996-04-01
影响因子：
4.1
通讯作者：
Lingelbach, K
中科院分区：
文献类型：
--
作者：
Ansorge, I;Benting, J;Lingelbach, K
恶性疟原虫是人类红细胞 (RBC) 的细胞内寄生虫。与许多其他细胞内寄生虫一样，恶性疟原虫在寄生液泡内驻留和发育，该寄生液泡由将宿主细胞的细胞质与寄生虫表面分开的膜结合。一些寄生虫蛋白被分泌到液泡空间中，而另一些则通过尚未明确的途径分泌到红细胞胞浆中。主要使用形态学方法跟踪寄生虫的蛋白质转运，为了寻找一个实验系统，该系统可以（i）剖析从寄生虫表面转运到红细胞胞浆中所涉及的各个步骤，以及（ii）评估液泡膜红细胞侧这一过程的分子需求，我们使用成孔蛋白链球菌溶血素 O 在保持液泡完整的条件下透化受感染的红细胞。对分别作为液泡空间和红细胞胞浆标记的两种寄生虫蛋白的分布进行形态学和生化分析。在透化红细胞中，两种标记蛋白被分选到与完整红细胞中相同的区室中。运往红细胞胞浆的蛋白质在跨过液泡膜之前穿过液泡空间。通过去除红细胞胞质，可以在液泡中阻止蛋白质运输。穿过液泡膜的易位需要 ATP 和膜红细胞表面的蛋白质来源，但它独立于红细胞的细胞内离子环境。
Plasmodium falciparum is an intracellular parasite of human red blood cells (RBCs). Like many other intracellular parasites, P. falciparum resides and develops within a parasitophorous vacuole which is bound by a membrane that separates the host cell cytoplasm from the parasite surface. Some parasite proteins are secreted into the vacuolar space and others are secreted, by an as yet poorly defined pathway, into the RBC cytosol. The transport of proteins from the parasite has been followed mainly using morphological methods, In search of an experimental system that would allow (i) dissection of the individual steps involved in transport from the parasite surface into the RBC cytosol, and (ii) an assessment of the molecular requirements for this process at the erythrocytic side of the vacuolar membrane, we have permeabilized infected RBCs with the pore-forming protein streptolysin O using conditions which left the vacuole intact. The distribution of two parasite proteins which served as markers for the vacuolar space and the RBC cytosol respectively was analysed morphologically and biochemically. In permeabilized RBCs the two marker proteins were sorted to the same compartments as in intact RBCs. The protein which was destined for the RBC cytosol traversed the vacuolar space before it was translocated across the vacuolar membrane. Protein transport could be arrested in the vacuole by removing the RBC cytosol. Translocation across the vacuolar membrane required ATP and a protein source at the erythrocytic face of the membrane, but it was independent of the intracellular ionic milieu of the RBC.