Phosphoproteome of Toxoplasma

弓形虫磷酸化蛋白质组

基本信息

批准号：
8224139
负责人：
John C Boothroyd
金额：
$ 23.46万
依托单位：
STANFORD UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-02-01 至 2014-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8224139
关键词：
Acute Affinity Chromatography Amino Acids Biochemical Bioinformatics Biological Blood capillaries Calcium Calcium Signaling Cell Culture Techniques Cells Communicable Diseases Coupled Data Data Set Development Disease Engineering Epitopes Event Fatal Outcome Fibroblasts Goals Human Human body Individual Infection Invaded Ion-Exchange Chromatography Procedure Ionophores Isotope Labeling Label Learning Link Location Malaria Malignant Neoplasms Maps Mass Spectrum Analysis Metals Methods Modeling Modification Molecular Organ Transplantation Organism Parasites Parasitic Diseases Patients Pharmaceutical Preparations Phosphopeptides Phosphorylation Phosphotransferases Plasmodium Process Protein Kinase Proteins Proteome RH1 Regulation Sampling Signal Pathway Signal Transduction Stable Isotope Labeling Staging System Techniques Technology Time Toxoplasma Toxoplasma gondii Toxoplasmosis Transplant Recipients Trypsin Work base calcium-dependent protein kinase capillary cell motility chemical genetics improved innovation liquid chromatography mass spectrometry mutant novel novel therapeutic intervention obligate intracellular parasite penis foreskin success

项目摘要

DESCRIPTION (provided by applicant): Apicomplexan parasites cause many diseases in humans, including malaria and toxoplasmosis. They do this by invading, replicating and exiting human cells. In recent years, much has been learned about the molecules that carry out these processes but how their actions are coordinated and how signals are received to initiate each process have remained largely mysterious. From studies in other organisms, it is known that phosphorylation of proteins is one of the most important means by which their action is regulated. Recently, mass spectrometric techniques have emerged that allow the large-scale identification of the nearly entire set of such modifications; this dataset is known as the "phosphoproteome". The goal of our proposed work is to determine the phosphoproteomic changes that occur in the acute stage of Toxoplasma parasites, the so-called tachyzoites, under intracellular vs. egressing conditions. To enable us to determine which changes are key to these processes, we will use specific drugs that disrupt calcium-dependent events that are known to be key to invasion and egress. We will also make strategic use of existing mutant lines of Toxoplasma that are defective in signaling related to invasion/egress. Finally, we will take advantage of our expertise in mass spectrometry to innovate new methods for phosphoproteomic analyses of samples where the amount available is too small for conventional analyses. PUBLIC HEALTH RELEVANCE: The work described here is relevant to the control of important parasitic diseases such as toxoplasmosis and malaria. Specifically, it will increase our understanding of the mechanisms used by these parasites to enter and exit cells. It also has the potential to improve our ability to study the molecular changes that occur in a variety of conditions, including other infections and cancer.

描述（由申请人提供）：顶复门寄生虫会引起人类许多疾病，包括疟疾和弓形体病。它们通过侵入、复制和退出人体细胞来做到这一点。近年来，人们对执行这些过程的分子有了很多了解，但它们的行为如何协调以及如何接收信号以启动每个过程仍然很大程度上是个谜。通过对其他生物体的研究，我们知道蛋白质的磷酸化是调节其作用的最重要手段之一。最近，出现了质谱技术，可以大规模鉴定几乎整组此类修饰；该数据集被称为“磷酸化蛋白质组”。我们提出的工作的目标是确定弓形虫寄生虫（所谓的速殖子）在细胞内与排出条件下的急性阶段发生的磷酸蛋白质组变化。为了使我们能够确定哪些变化是这些过程的关键，我们将使用特定的药物来破坏已知对入侵和流出至关重要的钙依赖性事件。我们还将战略性地利用现有的弓形虫突变株系，这些突变株系在与入侵/外出相关的信号传导方面存在缺陷。最后，我们将利用我们在质谱方面的专业知识，创新对样品进行磷酸化蛋白质组分析的新方法，其中可用的量对于传统分析来说太少。公共卫生相关性：此处描述的工作与弓形体病和疟疾等重要寄生虫病的控制相关。具体来说，它将增加我们对这些寄生虫进出细胞的机制的理解。它还有可能提高我们研究各种情况下发生的分子变化的能力，包括其他感染和癌症。