Zinc Trafficking Machinery in Malaria Infection

疟疾感染中的锌贩运机器

• 批准号: 7492154
• 负责人: Heather Relyea McGinley
• 金额: $ 0.71万
• 依托单位: NORTHWESTERN UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-06-01 至 2008-06-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7492154
• 关键词: Accounting; Affinity; Antibodies; Antimalarials; Area; Binding; Biochemistry; Bioinformatics; Biological Assay; Calorimetry; Carrier Proteins; Cell membrane; Cells; Cessation of life; Characteristics; Clinical; Collaborations; Communicable Diseases; Condition; Copper; Cytoplasm; Data; Depth; Detection; Development; Disease; Drug resistance; Environment; Erythrocyte Membrane; Erythrocytes; Fluorescence; Fluorescence Microscopy; Fluorescent Probes; Gene Expression Profile; Genes; Genetic Transcription; Genome; Genomics; Green Fluorescent Proteins; Growth; Homologous Gene; Human; Immunofluorescence Immunologic; In Vitro; Infection; Institutes; Ions; Iron; Knock-out; Knowledge; Lead; Life; Life Cycle Stages; Literature; Localized; Location; Malaria; Manuscripts; Measures; Membrane; Messenger RNA; Metals; Methods; Microarray Analysis; Monitor; Parasites; Pharmacologic Substance; Physiology; Plasmodium; Plasmodium falciparum; Population; Preparation; Proliferating; Protein Analysis; Proteins; Recombinant Proteins; Recruitment Activity; Relative (related person); Reporting; Resistance; Resolution; Roentgen Rays; Role; Saccharomyces cerevisiae; Staging; Starvation; Stress; Techniques; Technology; Testing; Time; Universities; Variant; Work; X ray microscopy; Zinc; base; divalent metal; drug sensitivity; extracellular; genetic manipulation; information gathering; intracellular protein transport; medical schools; professor; protein function; protein localization location; protein transport; sensor; trafficking; uptake; zinc-binding protein

DESCRIPTION (provided by applicant): Project Summary: All living cells must recruit metal ions to meet a certain quota, and the necessary concentration of many essential metals, including zinc, is relatively conserved among divergent species. However, in human erythrocytes parasitemized by the malaria parasite Plasmodium falciparum, zinc is found to accumulate in localized areas to a concentration of up to 20 times that of a normal erythrocyte. Interestingly, the intracellular levels of other essential metals such as copper and iron are not significantly altered by the presence of the parasite. In this proposal, we outline an approach to studying the mechanism by which the malaria parasite is able to dramatically alter the host cell zinc physiology. We believe that the proteins involved in zinc scavenging and hoarding serve as potential pharmaceutical targets in the effort to control this deadly parasite. This project will provide fundamental knowledge about this infectious disease and allow for the development of new treatments which will help to circumvent the drug resistances that currently hamper eradication of the disease. We aim to study the metallobiochemistry of candidate zinc import and transport proteins to develop an in- depth understanding of how the intracellular malaria parasite can manipulate the zinc quota in order to infect and proliferate in the host. Specifically, we aim to (1) identify candidate proteins by studying P. falciparum transcription profiles under zinc stress conditions, (2) examine the roles of these and known transporter homologs in subcellularzinc localization, and (3) examine the subcellular localization of the proteins themselves as well as their ability to bind and transport zinc in vitro. The methods used to accomplish these aims will include DMA microarray analysis of mRNA, determination of total and local zinc content in variant strains using X-ray fluorescence and fluorescent zinc sensors, and detection of protein location via immunofluorescent assays. Relevance: It is estimated that nearly 10% of the world's population will suffer from clinical malaria each year, resulting in between 1.5 and 2.7 million deaths annually. One characteristic that sets red blood cells infected with the malaria parasite apart from healthy red blood cells is the presence of elevated levels of zinc. Therefore we believe that studying the role of zinc in malaria proliferation will lead to a better understanding of this disease as well as suggesting new types of anti-malarial therapies.

项目概述：所有活细胞都必须补充金属离子以满足一定的配额，许多必需金属（包括锌）的必要浓度在不同物种中相对保守。然而，在被疟疾寄生虫恶性疟原虫寄生的人类红细胞中，发现锌在局部区域积聚，浓度高达正常红细胞的20倍。有趣的是，其他必需金属如铜和铁的细胞内水平不会因寄生虫的存在而发生显着改变。在这个建议中，我们概述了一种方法来研究疟疾寄生虫能够显着改变宿主细胞锌生理的机制。我们认为，参与锌清除和囤积的蛋白质是控制这种致命寄生虫的潜在药物靶点。该项目将提供有关这种传染病的基本知识，并允许开发新的治疗方法，这将有助于规避目前阻碍根除这种疾病的耐药性。我们的目标是研究候选锌输入和转运蛋白的金属生物化学，以深入了解细胞内疟原虫如何操纵锌配额，以便在宿主中感染和增殖。具体来说，我们的目标是（1）通过研究锌胁迫条件下恶性疟原虫的转录谱来鉴定候选蛋白，（2）检查这些蛋白和已知的转运蛋白同源物在亚细胞锌定位中的作用，（3）检查蛋白本身的亚细胞定位以及它们在体外结合和转运锌的能力。用于实现这些目标的方法将包括mRNA的DMA微阵列分析，使用X射线荧光和荧光锌传感器测定变异株中的总锌和局部锌含量，以及通过免疫荧光测定检测蛋白质位置。相关性：据估计，每年将有近10%的世界人口患有临床疟疾，每年导致150万至270万人死亡。感染疟原虫的红细胞与健康红细胞的一个特征是锌水平升高。因此，我们认为，研究锌在疟疾扩散中的作用将有助于更好地了解这种疾病，并提出新型抗疟疾疗法。