New Amino Acids for Protein Engineering

用于蛋白质工程的新氨基酸

• 批准号: 8134242
• 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
• 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/8134242
• 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

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复杂细胞系统中蛋白质何时何地产生。反应性氨基酸类似物将被掺入细胞蛋白质中，并选择性地与探针缀合，用于原核和真核细胞中新合成蛋白质的可视化、分离和鉴定。最重要的是，这些方法可以在细胞蛋白质合成分析中提供空间和时间分辨率。细胞选择性是通过突变氨酰-tRNA 合成酶的受控表达来实现的；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处于活性状态的细胞。该项目将探索使用此类方法来阐明细菌逃避哺乳动物宿主防御的机制，检查群体感应过程（这对于细菌病原体表达毒力至关重要），并以细胞选择性方式探究模型生物秀丽隐杆线虫中的蛋白质合成。这些研究将为从发育到疾病治疗的生物现象的系统级表征建立强大的通用平台。 公共健康相关性：该项目将提供新方法来阐明细菌试图逃避哺乳动物宿主防御的机制，检查细菌如何相互沟通以表达毒力因子并形成抗生素抗性生物膜，并识别活体动物中单个细胞产生的不同蛋白质组。这些研究将为从生长发育到传染病治疗的生物现象建立新的窗口。