ROLE OF CHAPERONES IN MAINTAINING THE ASPARAGINE REPEAT-RICH PROTEOME OF P. FALCI

伴侣在维持 P. FALCI 富含天冬酰胺重复蛋白质组中的作用

• 批准号: 8281043
• 负责人: Vasant Muralidharan
• 金额: $ 3.69万
• 依托单位: WASHINGTON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-05-01 至 2012-10-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8281043
• 关键词: Affect; Affinity; Affinity Chromatography; Amino Acids; Amyloid; Amyloid Fibrils; Animal Model; Anopheles Genus; Asparagine; Award; Binding; Biochemical; Biological; Biological Models; Biology; Biophysics; Cells; Cessation of life; Communicable Diseases; Complement; Complex; Culicidae; Development; Development Plans; Disease; Drug Delivery Systems; Drug resistance; Ensure; Exposure to; Falciparum Malaria; Fever; Foundations; Genes; Genomics; Glutamine; Guanine Nucleotide Exchange Factors; Heat shock proteins; Heat-Shock Response; Homologous Gene; Human; In Vitro; Insect Vectors; Institution; Knock-out; Lead; Life; Life Cycle Stages; Malaria; Mammalian Cell; Masks; Medicine; Mentors; Mentorship; Microscopic; Microscopy; Molecular Chaperones; Nucleotides; Organism; Paper; Parasites; Parasitology; Pharmaceutical Preparations; Phase; Phenotype; Plasmodium falciparum; Play; Process; Property; Protein Biochemistry; Proteins; Proteome; Reporting; Research; Research Personnel; Role; Scanning; Science; Specificity; Staging; Stress; System; Temperature; Text; Training; Universities; Washington; Yeasts; abstracting; amyloid fibril formation; amyloidogenesis; asexual; career; career development; extreme temperature; insight; killings; medical schools; member; multicatalytic endopeptidase complex; mutant; novel; pathogen; prevent; programs; protein function; protein misfolding; protein protein interaction

DESCRIPTION (provided by applicant): Abstract PI: MURALIDHARAN, VASANT Project: 1K99AI099156-01 Title: ROLE OF CHAPERONES IN MAINTAINING THE ASPARAGINE REPEAT-RICH PROTEOME OF P. FALCI Accession Number: 3400178 ================== NOTICE: THIS ABSTRACT WAS EXTRACTED FROM APPLICATION AND HAS NOT BEEN PROOFED BY AN SRA.WHEN THERE ARE PROBLEMS WITH THE APPLICATION SCANNING PROCESS, THE EXTRACTED TEXT MAY BE INCORRECT OR INCOMPLETE. ================== Plasmodium falciparum is a deadly human pathogen that is responsible for over a million deaths every year. The proteome of P. falciparum is disproportionately rich in repeats of asparagine; one in four proteins contain such repeats. The presence of Asn repeats in a protein leads to amyloid fibril formation. During its life cycle, P. falciparum undergoes large temperature fluctuations that can promote amyloid fibril formation. Chaperones or heat shock proteins, such as Hsp110, Hsp70 and Hsp40, are present in all kingdoms of life and influence amyloid fibril formation. However, their biological roles and mechanism of action remains poorly defined. The proposed career development plan aims to gain fundamental insights into the function of chaperones in maintaining the Asn repeat-rich proteome of P. falciparum, while establishing an independent academic career in a university setting. The candidate will build on his strong foundation in biochemistry, protein biophysics and parasitology to develop into an independent researcher in P. falciparum biology under the mentorship of Dr. Daniel Goldberg, a pioneer and leader in the study of the intraerythrocytic stages of the malaria parasite. The plan will be carrid out in the Department of Medicine, Division of Infectious Diseases at Washington University School of Medicine, a leading institution in biomedical science. In a recent paper, the candidate reported the development of a novel regulatable, fluorescent, affinity (RFA) tag that was used to study a 28-residue Asn repeat in the P. falciparum proteasome subunit, Rpn6. The Asn repeat did not affect the protein function, stability or interactions even under heat shock; suggesting th presence of an active process that ensures proper folding of Asn repeat containing proteins. During the mentored phase, the candidate will: 1) Examine the function of PfHsp110 in parasite proteostasis using the parasite lines where PfHsp110 gene has been RFA tagged via biochemical and microscopic approaches. Amyloid forming proteins will also be expressed in these parasite lines to study how the PfHsp110 influences fibril formation. 2) Determine the mechanism of action and specificity by complementing the RFA tagged parasite with mutants of PfHsp110 and homologs from other organisms. PfHsp110 will also be expressed in a heterologous mammalian system and its effect on fibril formation in mammalian cells will be studied using biochemical and microscopic approaches. In the independent phase of the award the candidate will elucidate the roles of Hsp70, Hsp70-like and Hsp40 chaperones of the parasite in maintaining a stable Asn repeat-rich proteome. These chaperones were discovered by the candidate to be associated with PfHsp110. Training in biochemical, microscopic and cell biological approaches as outlined in the research plan has equipped the candidate to embark on a comprehensive and fruitful research program as an independent researcher. The proposed studies will reveal fundamental aspects of chaperone function in P. falciparum, which will furnish new drug targets against malaria as well as other protein misfolding diseases. Relevance Malaria is a deadly disease that afflicts hundreds of millions of people worldwide. The parasite that causes the deadliest form of malaria, Plasmodium falciparum, is rapidly gaining drug resistance making it imperative to identify new strategies to kill this parasite. The proposed research will investigate new classes of drug targets to understand their role in parasite biology and develop better drugs against them.

描述（由申请人提供）： 抽象的 PI：Vasant项目Muralidharan：1K99AI099156-01标题：伴侣在维持芦笋重复富含P. Falci登录号的角色：3400178 =================== 注意：此摘要是从应用程序中提取的，尚未由SRA证明。当应用程序扫描过程存在问题时，提取的文本可能不正确或不完整。 =================== 恶性疟原虫是一种致命的人类病原体，每年造成超过100万人死亡。恶性疟原虫的蛋白质组富含天冬酰胺的重复。四分之一的蛋白质包含这种重复。蛋白质中ASN重复的存在导致淀粉样蛋白纤维形成。在其生命周期中，恶性疟原虫会经历较大的温度波动，可以促进淀粉样蛋白纤维形成。伴侣或热休克蛋白（例如HSP110，HSP70和HSP40）都存在于生命和影响淀粉样蛋白纤维形成的所有王国中。但是，它们的生物学作用和作用机制仍然很差。拟议的职业发展计划旨在获得对伴侣在维持ASN重复富含恶性疟原虫的重复蛋白质组功能的基本见解，同时在大学环境中建立独立的学术生涯。该候选人将基于他在生物化学，蛋白质生物物理学和寄生虫学领域的坚实基础，在丹尼尔·戈德堡（Daniel Goldberg）博士的指导下发展成为恶性疟原虫生物学的独立研究人员，后者是对马拉里亚寄生虫内部寄生虫的研究阶段的先驱和领导者的指导。 。该计划将在华盛顿大学医学学院的传染病科医学系（生物医学科学领域的领先机构）中进行。在最近的一篇论文中，候选人报道了一种新型可调节的，荧光，亲和力（RFA）标签的发展，该标签用于研究在恶性疟原虫蛋白酶体蛋白酶体亚基RPN6中重复28次。即使在热休克下，ASN重复也不会影响蛋白质功能，稳定性或相互作用。暗示存在一个活跃过程，该过程可确保正确折叠含有蛋白质的ASN重复。在指导阶段，候选人将：1）使用PFHSP110基因通过生化和显微镜方法对PFHSP110基因进行RFA的寄生虫线检查PFHSP110的功能。淀粉样蛋白形成蛋白也将在这些寄生虫线中表达，以研究PFHSP110如何影响原纤维形成。 2）通过与PFHSP110的突变体和其他生物体的同源物补充带有RFA标记的寄生虫来确定作用和特异性的机理。 PFHSP110还将在异源哺乳动物系统中表达，其对哺乳动物细胞中原纤维形成的影响将使用生化和微观方法研究。在奖励的独立阶段，候选人将阐明寄生虫的Hsp70，Hsp70样和HSP40伴侣的作用，以维持稳定的ASN重复蛋白质组。候选人发现这些伴侣与PFHSP110相关。研究计划中概述的生化，显微镜和细胞生物学方法的培训使候选人能够作为一项独立研究人员启动一项全面而富有成果的研究计划。拟议的研究将揭示恶性疟原虫中伴侣功能的基本方面，该方面将提供针对疟疾以及其他蛋白质错误折叠疾病的新药物靶标。相关性疟疾是一种致命的疾病，困扰着全球数亿人。导致最致命形式的疟疾疟原虫恶性疟原虫的寄生虫正在迅速获得耐药性，因此必须确定杀死这种寄生虫的新策略。拟议的研究将研究新的药物靶标，以了解其在寄生虫生物学中的作用，并针对它们开发更好的药物。