Erythrocyte Subversion by Malaria Parasite Exported Effectors

疟疾寄生虫输出效应子引起的红细胞颠覆

• 批准号: 9762189
• 负责人: Josh Ryan Beck
• 金额: $ 24.9万
• 依托单位: IOWA STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-08-15 至 2021-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9762189
• 关键词: ATP phosphohydrolase; Architecture; Biochemistry; Blood; CRISPR interference; CRISPR/Cas technology; Cause of Death; Cells; Cessation of life; Characteristics; Chimera organism; Complement; Complex; Computer Simulation; Cysteine; Development; Development Plans; Disease; Drug Design; Engineering; Ensure; Enzymes; Erythrocytes; Escherichia coli; Event; Gene Expression; Genes; Genetic; Genetic Screening; Goals; Heat shock proteins; Host Defense; Human; In Vitro; Infection; Invaded; Knock-out; Length; Malaria; Maps; Mass Spectrum Analysis; Mediating; Membrane; Modification; Molecular; Molecular Chaperones; Mutation; N-terminal; Nature; Paper; Parasites; Parasitic Diseases; Pathologic; Pathology; Pathway interactions; Peptide Hydrolases; Plasmodium; Plasmodium falciparum; Process; Property; Protein Export Pathway; Proteins; Proteomics; Recombinants; Regulation; Reporter; Research; Research Personnel; Resolution; Role; Structural Protein; Structure; Substrate Specificity; System; TXN gene; Testing; Therapeutic; Vacuole; Virulence; Work; aptamer; base; career; career development; crosslink; experience; experimental study; forward genetics; knock-down; mutant; new therapeutic target; novel; novel therapeutics; prevent; protein E; protein function; protein structure; protein transport; reverse genetics; therapeutic target; tool

Project Summary/Abstract Malaria remains one of the most devastating parasitic diseases in the world with the vast majority of deaths caused by Plasmodium falciparum. The pathology of the disease results exclusively from the blood-stage of the infection during which parasites invade and multiply within host erythrocytes. To establish this intracellular niche, P. falciparum imposes striking modifications to the erythrocyte through export of effector proteins but the export mechanism is poorly understood and the effector functions essential to parasite survival remain largely unknown. Export into the red blood cell requires crossing a vacuole membrane surrounding the parasite, a translocation event that depends upon the Plasmodium translocon of exported proteins (PTEX). In a recent paper, the candidate showed that inactivation of heat shock protein 101 (HSP101), a AAA+ ATPase component of PTEX, results in a complete block in protein export and parasite death. Related AAA+ proteins can unfold and directionally thread substrates through a central channel, suggesting HSP101 may drive recognition of effector proteins and power the translocation process. The proposed career development plan aims to dissect the role of HSP101/PTEX in export and to identify key exported effectors that enable the parasite to survive within the erythrocyte. Specific Aim 1 seeks to understand the substrate recognition and catalytic properties of recombinant HSP101 and to dissect the role of HSP101 in protein export within intact parasites. Specific Aim 2 will use genetic and proteomic approaches to define the function of additional PTEX components and a cross-linking mass spectrometry approach to map the architecture of the complex. Specific Aim 3 will analyze the function of exported effectors implicated in parasite survival within the erythrocyte using forward and reverse genetics as well as proteomic approaches. PTEX is an exciting new drug target and the proposed experiments will reveal key mechanistic information about the role of this export machinery as a necessary basis for rational drug design. Furthermore, identification of key exported proteins with roles in parasite survival may provide needed additional therapeutic targets. Collectively, this work will further our understanding of how the malaria parasite subverts its erythrocyte host cell and support development of new tools for control of malaria disease. The experience and research tools acquired in the process will propel the candidate into an established career as an independent investigator.

项目总结/摘要 疟疾仍然是世界上最具破坏性的寄生虫病之一， 大多数死亡是由恶性疟原虫引起的。疾病的病理结果 仅从血液阶段的感染，在此期间寄生虫入侵和繁殖 在宿主红细胞内。为了建立这种细胞内的生态位，恶性疟原虫施加了惊人的 通过输出效应蛋白修饰红细胞，但输出机制是 对寄生虫生存所必需的效应器功能仍然很大程度上 未知输出到红细胞需要穿过围绕在红细胞周围的空泡膜。 寄生虫，一个易位事件，取决于疟原虫易位输出 蛋白质（PTEX）。在最近的一篇论文中，候选人表明，热休克蛋白的失活 101（HSP 101），PTEX的AAA+ ATP酶组分，导致蛋白质的完全阻断， 输出和寄生虫死亡。相关的AAA+蛋白可以解折叠和定向穿线底物 通过中央通道，表明HSP 101可能驱动效应蛋白的识别， 为转移过程提供动力拟议的职业发展计划旨在剖析 HSP 101/PTEX在输出中的作用，并确定使寄生虫能够 在红细胞内存活。具体目标1旨在了解底物识别， 重组HSP 101的催化特性，并剖析HSP 101在蛋白输出中的作用 完整的寄生虫体内具体目标2将使用遗传和蛋白质组学方法来定义 额外的PTEX组分的功能和交联质谱法， 绘制建筑群的结构图具体目标3将分析导出的功能 使用正向和反向的红细胞内寄生虫存活的相关效应物 遗传学和蛋白质组学方法。PTEX是一个令人兴奋的新药物靶点， 所提出的实验将揭示关于这种输出的作用的关键机制信息 机械作为合理药物设计的必要基础。此外，识别密钥 在寄生虫存活中起作用的输出蛋白质可以提供所需的额外治疗 目标的总的来说，这项工作将进一步加深我们对疟疾寄生虫 破坏其红细胞宿主细胞，并支持开发控制疟疾的新工具 疾病在此过程中获得的经验和研究工具将推动候选人 成为一名独立调查员

项目成果

The PTEX Pore Component EXP2 Is Important for Intrahepatic Development during the Plasmodium Liver Stage.

• DOI: 10.1128/mbio.03096-22
• 发表时间: 2022-12-20
• 期刊: mBio
• 影响因子: 6.4