Diversity Supplement for Elucidating the trafficking mechanisms of effector proteins to the Plasmodium infected red blood cell

用于阐明效应蛋白向疟原虫感染的红细胞运输机制的多样性补充

• 批准号: 10077624
• 负责人: Vasant Muralidharan
• 金额: $ 6.42万
• 依托单位: UNIVERSITY OF GEORGIA
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-01-05 至 2022-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10077624
• 关键词: Affect; Antimalarials; Asparagine; Binding; Biochemical; Biological Assay; Biological Models; Biological Process; Biotin; Biotinylation; Blood Vessels; Blood capillaries; Brain; CRISPR/Cas technology; Catalytic Domain; Cell Communication; Cell membrane; Cells; Cellular Membrane; Cessation of life; Clinical; Coagulation Process; Complement; Complex; Cytoplasm; Development; Disease; Drug Targeting; Drug resistance; Endoplasmic Reticulum; Endothelial Cells; Erythrocytes; Eukaryota; Event; Flow Cytometry; Genes; Genetic; Genome; Glycine decarboxylase; Glycoproteins; Goals; Growth; Habitats; Home environment; Human; Immunofluorescence Immunologic; Infection; Label; Lead; Life Cycle Stages; Ligase; Link; Malaria; Malaria Vaccines; Membrane; Metabolism; Methods; Modification; Molecular; Movement; Mutation; Nutrient; Oligosaccharides; Orthologous Gene; Parasites; Parasitic Diseases; Pathway interactions; Permeability; Plasmodium; Plasmodium falciparum; Plasmodium falciparum genome; Polysaccharides; Process; Property; Protein Export Pathway; Protein Glycosylation; Proteins; Proteome; Research; Resistance; Resistance development; Role; Side; Site; Sum; Testing; Time; Usher Proteins; asexual; base; conditional mutant; data acquisition; drug development; experimental study; glycosylation; human disease; mutant; novel therapeutics; obligate intracellular parasite; parent grant; programs; protein function; protein transport; resistant strain; screening; trafficking; whole genome

RESEARCH SUMMARY [PARENT GRANT] Plasmodium falciparum is a deadly human parasite that causes malaria and is responsible for nearly 450,000 deaths every year. Malaria is endemic in large regions of the world, home to about 4 billion people and it affects ~250 million people annually. There are no effective vaccines against malaria and antimalarial drugs are the mainstay of treatment. At this time, the parasite has gained resistance to all clinically available antimalarial drugs and these drug resistant strains are spreading throughout the world, threatening all the progress that has been made against this disease in the last decade. Therefore, it is imperative that we constantly generate new drugs and identify potential drug targets to stay ahead of this nefarious disease. The clinical manifestations of this devastating parasitic disease, including death, are caused by the growth of P. falciparum within the host red blood cell (RBC). To build a suitable habitat for growth inside RBCs, the malaria parasite completely transforms the host cell. It changes the metabolism of the RBC, makes the RBC more rigid such that it is harder for the infected RBC to pass through capillaries, modifies the RBC membrane to allow for favorable movement of nutrients, and alters the binding properties of the RBC so that the infected cell can bind to the endothelial cells lining blood vessels. The sum of these changes leads to disease and death, for instance, binding of the P. falciparum infected RBC to endothelial cells can clog blood vessels in the brain leading to clots that eventually result in death. The subjugation of the infected RBC is accomplished through the action of several hundred proteins that the parasite transports to the host cell via poorly understood mechanisms. The export of parasite effector proteins is essential for transforming the RBC and therefore, for causing disease. Parasite effector proteins that are synthesized in the parasite cytoplasm need to be transported across three or four cellular membranes in order to reach their site of action in the host RBC. The molecular mechanisms that recognize, sort, and transport these parasite effectors to the infected RBC remain to be identified. The proposed studies aim to unravel the molecular processes that govern key early events that set parasite effectors on the path to the host RBC. We will pursue two aims to accomplish this goal. First, we will generate conditional mutants of proteins in the endoplasmic reticulum of the parasite that are potentially required for export of parasite effectors. The mutants will be analyzed using genetic, cellular, and biochemical approaches to determine their roles in the export of parasite proteins. Second, we will take an unbiased interactome screening approach that uses a proximity-based labeling approach and discover proteins that usher exported proteins to their site of action in the host RBC. Attaining the objectives of the research program will reveal key and unique protein trafficking mechanisms of P. falciparum that may be targeted for antimalarial drug development.

研究摘要[父母赠款] 恶性疟原虫是一种致命的人寄生虫，可引起疟疾，并造成近45万 每年死亡。疟疾在世界上大型地区是地方性的，有大约40亿人口，这会影响 每年约2.5亿人。没有针对疟疾的有效疫苗，抗疟药是 治疗中的支柱。目前，寄生虫对所有临床上可用的抗疟药都具有抵抗力 这些抗药性菌株正在全世界蔓延，威胁着所有的进步 在过去的十年中针对这种疾病。因此，我们必须不断生成新药 并确定潜在的药物靶标，以保持这种邪恶疾病的领先地位。临床表现 毁灭性的寄生疾病（包括死亡）是由恶性疟原虫在宿主红色内的生长引起的 血细胞（RBC）。为了建立一个适合RBC内生长的栖息地，疟疾寄生虫完全转变 宿主单元。它改变了RBC的代谢，使RBC更加僵化，因此很难 感染的RBC通过毛细血管，修饰RBC膜以进行有利的运动 营养，并改变RBC的结合特性，使感染细胞可以与内皮细胞结合 衬里血管。这些变化的总和导致疾病和死亡，例如，P的结合。 恶性疾病感染的RBC对内皮细胞会堵塞大脑中的血管，导致凝块最终 导致死亡。受感染的RBC的征服是通过数百个行动来完成的 寄生虫通过较知的机制传递到宿主细胞的蛋白质。寄生虫的出口 效应蛋白对于转化RBC，因此对于引起疾病至关重要。寄生虫效应子 在寄生虫细胞质中合成的蛋白质需要在三个或四个细胞上转运 膜以在主机RBC中到达其行动地点。识别的分子机制 排序并将这些寄生虫效应子运输到受感染的RBC上。提出的研究 旨在阐明主导关键早期事件的分子过程，这些事件将寄生虫效应子设置在通往道路的道路上 主机RBC。我们将追求两个目标来实现这一目标。首先，我们将产生有条件的突变体 寄生虫的寄生虫内质网中的蛋白质可能需要导出寄生虫效应子。 将使用遗传，细胞和生化方法对突变体进行分析，以确定它们在 寄生虫蛋白的出口。其次，我们将采用一种使用公正的互动筛选方法 基于接近性的标签方法并发现蛋白质将蛋白质引入其作用部位的蛋白质 主机RBC。达到研究计划的目标将揭示关键和独特的蛋白质运输 恶性疟原虫的机制可能针对抗疟药的针对性。