Phosphoproteome of Toxoplasma

弓形虫磷酸化蛋白质组

基本信息

批准号：
8416302
负责人：
John C Boothroyd
金额：
$ 19.55万
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/8416302
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.

描述（由申请人提供）：Apicomplexan寄生虫在包括疟疾和弓形虫病在内的人类中引起许多疾病。他们通过入侵，复制和退出人类细胞来做到这一点。近年来，已经了解了执行这些过程的分子，但是如何协调其行动以及如何收到启动每个过程的信号仍然很神秘。从其他生物体的研究中，众所周知，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，允许大规模识别几乎整个此类修改。该数据集被称为“磷蛋白组”。我们提出的工作的目的是确定在弓形虫寄生虫的急性阶段发生的磷酸蛋白质组学变化，即所谓的tachyzoites，在细胞内与出口条件下。为了使我们能够确定哪些变化是这些过程的关键，我们将使用破坏钙依赖性事件的特定药物，而钙依赖性事件已知是入侵和出口的关键。我们还将战略使用与入侵/出口有关的信号传导有缺陷的现有弓形虫突变型线。最后，我们将利用我们在质谱方面的专业知识来创新新方法，用于对可用量对于常规分析的样品的磷光蛋白质组学分析。