PfVP1, a battery of the malaria parasite, is critical for ring stage development

PfVP1 是疟疾寄生虫的一组，对于环期发育至关重要

• 批准号: 10116730
• 负责人: Hangjun Ke
• 金额: $ 22.56万
• 依托单位: DREXEL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-11-10 至 2022-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10116730
• 关键词: Affinity; Antimalarials; Archaea; Bacteria; Biochemical; Bioenergetics; Biological; Biology; Blood; Budgets; CRISPR/Cas technology; Cell membrane; Cells; Complement; Consumption; Contracts; DNA biosynthesis; Data; Development; Diphosphates; Drug resistance; Energy Supply; Energy-Generating Resources; Erythrocytes; Future; Genetic; Glucose; Glycolysis; Human; Hydrolysis; Knock-out; Malaria; Mammalian Cell; Measures; Mediating; Membrane; Metabolic; Molecular Target; Mutagenesis; Mutate; Nutrient; Organism; Oxidative Phosphorylation; Parasites; Physiological; Plants; Plasmodium falciparum; Process; Proteins; Proton Pump; Protons; RNA; Reaction; Regulatory Element; Resistance; Resistance development; Risk; Role; Route; Running; Saccharomyces cerevisiae; Testing; Time; Vacuole; Vesicle; Waste Products; Yeasts; asexual; base; cost; drug development; fungus; genomic locus; global health; insight; knock-down; metabolic rate; nutrition; pyrophosphatase; sodium ion; uptake

Project Summary Malaria remains a major global health concern and the control progress has run into a serious bottleneck in recent years. Each year, more than 3 billion people are living at risk of contracting malaria, 200 million people are infected and half a million people die from malaria. As malaria parasites rapidly develop resistance to available antimalarial drugs, a better understanding of fundamental biology is urgently needed. In the asexual blood stages, malaria parasites mainly reply on glycolysis for energy since the parasite's mitochondrion does not perform oxidative phosphorylation to produce ATP. They also contain membrane bound proton pumping pyrophosphatases (mPPases), which use pyrophosphate, the by-product of over 200 cellular reactions, as the energy source to pump protons across a membrane. We have previously knocked out PfVP2 (PF3D7_1235200) whereas PfVP1 (PF3D7_1456800) has been predicted to be essential. Using a CRISPR/Cas9 mediated genetic tagging/conditional knockdown, we have found that PfVP1 is localized to the parasite plasma membrane and is essential for ring stage development. In this application, we hypothesize that PfVP1 uses pyrophosphate as an alternative energy source to sustain a proton electrochemical gradient in the ring stage development of Plasmodium falciparum. We will test this hypothesis in the following aims. Aim 1, Assess the activity of PfVP1 as an integral membrane proton-pumping pyrophosphatase. Aim 2, Define the function of PfVP1 in ring stage parasites. Together, this proposal will uncover important insights of ring stage development and bioenergetics, critical yet understudied aspects of parasite biology. If successful, the studies proposed here will provide direct evidence that an alternative energy source is critical for early stage development of a malaria parasite in RBCs. Finding a critical molecular target at the ring stage will also facilitate target-based drug development in future to better inhibit metabolically less active parasite forms that are often resistant to current antimalarials.

项目摘要 疟疾仍然是一个主要的全球健康问题，控制进展遇到了严重的问题， 近年来的瓶颈。每年有超过30亿人生活在感染的风险中， 有2亿人感染疟疾，50万人死于疟疾。疟疾 寄生虫迅速对现有的抗疟药物产生耐药性， 基础生物学是迫切需要的。在无性血液阶段，疟原虫主要 回答糖酵解的能量，因为寄生虫的寄生虫不进行氧化 磷酸化产生ATP。它们还包含膜结合质子泵 焦磷酸酶（mPPases），它使用焦磷酸盐，超过200个细胞的副产物， 反应，作为能量源泵质子穿过膜。我们先前已经 敲除PfVP 2（PF3D7_1235200），而预测PfVP 1（PF3D7_1456800） 是必不可少的。使用CRISPR/Cas9介导的遗传标记/条件性敲低， 已经发现PfVP 1定位于寄生虫质膜，并且是环 阶段发展。在本应用中，我们假设PfVP 1使用焦磷酸盐作为 替代能源以维持环级中的质子电化学梯度 恶性疟原虫的发展。我们将在以下目标中检验这一假设。 目的1、研究PfVP 1作为膜质子泵焦磷酸酶的活性。 目的2、明确PfVP 1在环期寄生虫中的功能。这项提案将揭示 环阶段发展和生物能量学的重要见解，关键但未充分研究 寄生虫生物学的各个方面。如果成功，这里提出的研究将提供直接证据， 替代能源对于疟疾寄生虫的早期发展至关重要， 红细胞。在环阶段找到关键的分子靶点也将有助于靶向药物的开发。 未来的发展，以更好地抑制代谢活性较低的寄生虫形式， 对目前的抗疟药有抗药性。