Desai S A, Krogstad D J, McCleskey E W
Department of Cell Biology, Washington University, St Louis, Missouri 63110.
Nature. 1993 Apr 15;362(6421):643-6. doi: 10.1038/362643a0.
During its 48-hour cycle inside the red blood cell, the human malaria parasite, Plasmodium falciparum, increases its volume 25-fold and divides asexually. This rapid growth demands large amounts of nutrients, a problem exacerbated by the lower metabolic rate and relative ionic impermeability of the host red blood cell. Direct passage of small nutrients across the two membranes that separate the parasite from the erythrocyte cytosol may be important for parasite development and has been demonstrated for radiolabelled glucose, amino acids, and purine nucleosides. Flux studies on plasmodia are limited, however, to suspensions of erythrocyte-free parasites and so cannot be used to examine the individual transport properties of the two membranes involved. Here we use the cell-attached patch clamp method to overcome this limitation. After removing the intervening red blood cell membrane and forming gigaohm seals on the small (3-5 microns) parasite, we studied transport across the parasitophorous vacuole membrane (PVM), the outer of the two membranes that separate the parasite from the erythrocyte cytosol. A 140-pS channel which is permeable to both cations and anions was identified on the PVM. This channel is present at high density, is open more than 98 per cent of the time at the resting potential of the PVM, and is permeable to lysine and glucuronate. The channel can readily transport amino acids and monosaccharides across the PVM and may be essential for fulfilling the parasite's metabolic demands.
在人类疟原虫恶性疟原虫于红细胞内的48小时周期中,其体积增大25倍并进行无性分裂。这种快速生长需要大量营养物质,而宿主红细胞较低的代谢率和相对的离子不可渗透性使这一问题更加严重。小营养物质直接穿过将寄生虫与红细胞胞质溶胶分隔开的两层膜,这对寄生虫的发育可能很重要,并且已通过放射性标记的葡萄糖、氨基酸和嘌呤核苷得到证实。然而,对疟原虫的通量研究仅限于无红细胞寄生虫的悬浮液,因此不能用于检查所涉及的两层膜的个体转运特性。在这里,我们使用细胞贴附式膜片钳方法来克服这一限制。在去除中间的红细胞膜并在小(3 - 5微米)的寄生虫上形成千兆欧密封后,我们研究了穿过寄生泡膜(PVM)的转运,PVM是将寄生虫与红细胞胞质溶胶分隔开的两层膜中的外层膜。在PVM上鉴定出一个对阳离子和阴离子都通透的140皮安通道。该通道高密度存在,在PVM的静息电位下超过98%的时间处于开放状态,并且对赖氨酸和葡萄糖醛酸通透。该通道能够轻易地将氨基酸和单糖转运穿过PVM,可能对满足寄生虫的代谢需求至关重要。