EXPRESSION OF THE PLASMODIAL NUTRIENT CHANNEL ON OOCYTES

卵母细胞上疟原虫营养通道的表达

• 批准号: 2057456
• 负责人: SANJAY A DESAI
• 金额: $ 6.43万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-08-01 至 1997-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2057456
• 关键词: Plasmodium falciparum; Xenopus oocyte; adenosine triphosphate; antimalarial agents; erythrocyte membrane; erythrocytes; gene expression; human tissue; lactates; lipid bilayer membrane; membrane activity; membrane channels; membrane permeability; membrane reconstitution /synthesis; membrane transport proteins; messenger RNA; microinjections; nutrient intake activity; nutrition related tag; parasitism; potassium channel; second messengers; tissue /cell culture; vesicle /vacuole; voltage /patch clamp

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.

这是分离和表征营养渗透性离子的提议 红细胞内疟疾寄生虫恶性疟原虫的通道。 这 通道首先由申请人和他的同事描述[1， 附录]并且可能对于寄生虫的生存至关重要，因为它允许 寄生虫从红血中获取氨基酸和单糖 细胞（红细胞）。 该通道是通过使用膜片钳方法测量离子来识别的 寄生液泡膜 (PVM) 上的通道活性 寄生虫周围有两层膜。 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，表达的通道将用于测量 K+、乳酸和 ATP4- 渗透性，检查通道调节，并调查可能的情况 阻滞剂。 如果在卵母细胞上表达不成功，则采用替代方法 方法，重构为脂质双层，将允许相同的细节 表征。 这些研究应该解决以下基本问题： 寄生虫营养物质贩运，可能对 疟疾治疗：一种已确定的通道阻断剂可能会阻止营养 获取并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。 他有相当多的 具有用于研究膜运输和意志的方法的经验， 该奖项为分子生物学方法奠定了基础。