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: 2057455
• 负责人: SANJAY A DESAI
• 金额: $ 5.93万
• 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/2057455
• 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.

这是隔离和表征营养可渗透离子的建议 恶性疟原虫肠内疟疾寄生虫的通道。 这 渠道最初是由申请人及其同事[1， 附录]，对于寄生虫生存可能是必不可少的，因为它允许 从红血中获取氨基酸和单糖的寄生虫 细胞（RBC）。 通过使用斑块夹方法测量离子，可以识别此通道 寄生虫液泡膜（PVM）上的通道活动 寄生虫周围的两个膜。 申请人现在建议 通过对寄生虫的微注射在爪蟾卵母细胞上表达此通道 mRNA。 然后，表示的通道将用于测量K+，乳酸和 ATP4-渗透率，检查通道调节并研究可能 阻滞剂。 如果在卵母细胞上的表达不成功，则替代 方法，重建为脂质双层，将允许相同的详细信息 表征。 这些研究应解决有关的基本问题 寄生虫的营养贩运，可能对 疟疾的治疗：已识别的通道阻滞剂可能会防止营养 因此，通过饥饿杀死寄生虫。 申请人已决定将他的长期研究工作集中在 RBC内质量存活的机制。 他有很大的 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。