EXPRESSION OF THE PLASMODIAL NUTRIENT CHANNEL ON OOCYTES

这是一个建议，分离和表征营养渗透离子 红细胞内疟疾寄生虫，恶性疟原虫。 这 信道首先由申请人及其同事描述[1， 阑尾]，可能是寄生虫生存所必需的，因为它允许 寄生虫从红血中获取氨基酸和单糖 红细胞（RBC）。 用膜片钳方法测定了该通道的离子通道 通道活动的寄生虫空泡膜（PVM），外 寄生虫周围的两层膜。 申请人现建议 通过显微注射寄生虫在非洲爪蟾卵母细胞上表达该通道 mRNA。 然后将使用表达的通道测量K+、乳酸和 ATP 4渗透性，检查通道调节，并研究可能的 阻滞剂。 如果在卵母细胞上表达不成功， 方法，重建成脂质双层，将允许相同的详细说明， 特征化 这些研究应解决以下基本问题： 寄生虫的营养运输，并可能有深远的影响， 治疗疟疾：一种已确定的通道阻滞剂可能会阻止营养素 因此，通过饥饿杀死寄生虫。 申请人决定将其长期研究工作集中在 红细胞内疟原虫存活的机制。 他有相当大的 有研究膜转运方法的经验， 这个奖项，在分子生物学的方法发展的基础。

This is a proposal to isolate and characterize the nutrient-permeable ion channel on the intraerythrocytic malaria parasite, P. falciparum. This channel was first described by the applicant and his colleagues [1, appendix] and may be essential for parasite survival because it permits the parasite to acquire amino acids and monosaccharides from the red blood cell (RBC). This channel was identified by using the patch clamp method to measure ion channel activity on the parasitophorous vacuole membrane (PVM), the outer of two membranes surrounding the parasite. The applicant now proposes to express this channel on Xenopus oocytes by microinjection of parasite mRNA. Expressed channels will then be used to measure K+, lactate, and ATP4- permeabilities, examine channel regulation, and investigate possible blockers. If expression on oocytes is unsuccessful, an alternative approach, reconstitution into lipid bilayers, will allow the same detailed characterization. These studies should address fundamental questions on parasite nutrient trafficking and may have profound implications for the treatment of malaria: an identified channel blocker may prevent nutrient acquisition and, therefore, kill parasites through starvation. The applicant has decided to focus his long-term research efforts on the mechanisms of plasmodial survival within the RBC. He has considerable experience with methods used to study membrane transport and will, with this award, develop a foundation in the methods of molecular biology.