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

研究摘要(家长资助金) 恶性疟原虫是一种致命的人类寄生虫，可导致疟疾，造成近45万人死亡 每年都有人死亡。疟疾在世界上大片地区流行，约有40亿人口，它影响到 每年约有2.5亿人。目前还没有有效的疟疾疫苗，抗疟疾药物是 治疗中流砥柱。目前，这种寄生虫已经对所有临床上可用的抗疟疾药物产生了抗药性。 这些耐药菌株正在世界各地传播，威胁着所有已经取得的进展 在过去的十年里针对这种疾病所做的努力。因此，我们必须不断制造新的药物。 并确定潜在的药物靶点，以保持领先于这种邪恶的疾病。该病的临床表现 毁灭性的寄生虫病，包括死亡，是由恶性疟原虫在宿主红色体内的生长引起的。 血细胞(RBC)。为了在红细胞内建立适合生长的栖息地，疟疾寄生虫完全改变了 宿主细胞。它改变了红细胞的新陈代谢，使红细胞变得更加僵硬，从而使 感染的红细胞通过毛细血管，修改红细胞膜以允许有利的运动 营养物质，并改变红细胞的结合特性，使受感染的细胞可以与内皮细胞结合 血管衬里。这些变化的总和导致疾病和死亡，例如，P。 恶性疟原虫感染内皮细胞的红细胞会堵塞大脑中的血管，最终导致血栓 导致死亡。征服受感染的红细胞是通过数百人的行动完成的。 寄生虫通过鲜为人知的机制向宿主细胞运输的蛋白质。寄生虫的出口 效应蛋白是转化红细胞的关键，因此也是致病的关键。寄生虫效应器 在寄生虫细胞质中合成的蛋白质需要通过三到四个细胞运输 膜才能到达它们在宿主红细胞中的作用部位。识别的分子机制， 对这些寄生虫效应器进行分类，并将其运送到受感染的红细胞中仍有待鉴定。建议进行的研究 旨在揭开控制关键早期事件的分子过程，这些事件将寄生虫效应器设置在 宿主RBC。为实现这一目标，我们将追求两个目标。首先，我们将生成条件突变 寄生虫内质网中的蛋白质，可能是输出寄生虫效应器所必需的。 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，我们将采取无偏见的互动组筛选方法，使用 基于邻近度的标记方法和发现将输出蛋白质引入其作用部位的蛋白质 宿主RBC。实现研究计划的目标将揭示关键和独特的蛋白质交易 恶性疟原虫抗疟疾药物开发的作用机制。

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.