New Amino Acids for Protein Engineering

用于蛋白质工程的新氨基酸

基本信息

批准号：
8309176
负责人：
DAVID A TIRRELL
金额：
$ 35.3万
依托单位：
CALIFORNIA INSTITUTE OF TECHNOLOGY
依托单位国家：
美国
项目类别：
财政年份：
2001
资助国家：
美国
起止时间：
2001-01-01 至 2014-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8309176
关键词：
Amino Acids Amino Acyl-tRNA Synthetases Anatomy Animal Model Animals Antibiotic Resistance Bacteria Bacterial Proteins Behavior Biological Models Biological Phenomena Caenorhabditis elegans Cataloging Catalogs Cell Count Cell Culture Techniques Cells Complex Data Development Disease Dissection Drug Metabolic Detoxication Enhancers Epithelial Cells Escherichia coli Eukaryotic Cell Food Funding Gene Expression Genes Growth and Development function Hydrogen Peroxide Imagery Individual Intestines Label Life Ligase Mammalian Cell Mass Spectrum Analysis Measurement Measures Metabolic Methods Microbial Biofilms Muscle Cells Nematoda Nervous system structure Neurons Nutrient Organism Output Oxidative Stress Phagocytosis Process Production Prokaryotic Cells Protein Biosynthesis Protein Engineering Proteins Proteome Proteomics Regulon Resolution Set protein Stress Superoxides System Time Transgenic Organisms Translating Vibrio Virulence Virulence Factors analog biological adaptation to stress cell type genome sequencing in vivo infectious disease treatment macrophage microbial mutant pathogen pharynx muscle programs promoter public health relevance quorum sensing research study response tool transcription factor uptake

项目摘要

DESCRIPTION (provided by applicant): This project will develop powerful new tools to determine when and where proteins are made in complex cellular systems. Reactive amino acid analogs will be incorporated into cellular proteins and selectively conjugated to probes for visualization, isolation and identification of newly synthesized proteins in both prokaryotic and eukaryotic cells. Most importantly, these approaches can provide both spatial and temporal resolution in analysis of cellular protein synthesis. Cell-selectivity is achieved by controlled expression of mutant aminoacyl-tRNA synthetases; in systems containing multiple cell types, amino acid labeling is confined to cells in which the mutant synthetase is active. This project will explore the use of such methods to elucidate the mechanisms by which bacteria evade the defenses of mammalian hosts, to examine the process of quorum sensing (which is essential to the expression of virulence by bacterial pathogens), and to interrogate protein synthesis in a cell-selective manner in the model organism Caenorhabditis elegans. These studies will establish powerful, general platforms for systems-level characterization of biological phenomena ranging from development to the treatment of disease. PUBLIC HEALTH RELEVANCE: This project will provide new methods to elucidate the mechanisms by which bacteria attempt to evade the defenses of their mammalian hosts, to examine how bacteria communicate with one another to express virulence factors and form antibiotic-resistant biofilms, and to identify the different sets of proteins made by individual cells in living animals. These studies will establish new windows on biological phenomena ranging from growth and development to the treatment of infectious disease.

描述（由申请人提供）：该项目将开发强大的新工具来确定在复杂的蜂窝系统中何时何地制作蛋白质。反应性氨基酸类似物将掺入细胞蛋白中，并选择性地结合到探针中，以可视化，分离和鉴定原核细胞和真核细胞中新合成的蛋白质。最重要的是，这些方法可以在分析细胞蛋白合成时提供空间和时间分辨率。通过控制突变氨基酰基-TRNA合成酶的控制表达来实现细胞选择性。在包含多种细胞类型的系统中，氨基酸标记仅限于突变合成酶活性的细胞。该项目将探讨使用这种方法来阐明细菌逃避哺乳动物宿主的防御能力的机制，检查Quorum传感的过程（这对于细菌病原体表达毒力至关重要），以及在模型生物体caenenorhabditis elementiation的细胞中质量蛋白质合成的蛋白质合成。这些研究将建立强大的一般平台，用于系统级的生物学现象表征，从发育到治疗疾病。公共卫生相关性：该项目将提供新的方法来阐明细菌试图逃避其哺乳动物宿主的防御能力的机制，以研究细菌如何相互沟通以表达毒力并形成抗生素耐药性生物膜，并识别单个细胞在活动物中产生的蛋白质的不同集合。这些研究将建立有关从生长和发育到传染病治疗的生物学现象的新窗口。