Eukaryotic incorporation of photoreactive amino acids

光反应性氨基酸的真核掺入

• 批准号: 6878087
• 负责人: JAMES TURNER
• 金额: $ 4.13万
• 依托单位: SCRIPPS RESEARCH INSTITUTE, THE
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2006-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6878087
• 关键词: Caenorhabditis elegans; aging; aminoacid; cell surface receptors; crosslink; eukaryote; fluorescence microscopy; fluorescent dye /probe; green fluorescent proteins; ketones; mass spectrometry; phenylalanine; postdoctoral investigator; protein protein interaction; proteomics; reagent /indicator; site directed mutagenesis

DESCRIPTION (provided by applicant): Unnatural amino acids will be incorporated in vivo at specific positions within G-protein coupled receptors in order to fluorescently label proteins and detect and characterize protein/protein and protein/ligand interactions. Specific labeling of a single amino acid within a protein with a fluorescent label will provide information about the local microenvironment of that residue. Labeling with a photocrosslinking amino acid, followed by shotgun proteomics by the Biological Mass Spectrometry Lab at TSRI, will identify ligand interactions of that residue in human orphan receptors expressed in yeast. Photocrosslinking using an unnatural amino acid incorporated in C. elegans will be used to characterize a pathway involved with aging. Work with C. elegans will be in collaboration with Andrew Dillin at The Salk Institute.

描述（由申请人提供）： 不自然的氨基酸将在G蛋白偶联受体中的特定位置中掺入体内，以便荧光标记蛋白质并检测和表征蛋白质/蛋白质和蛋白质/配体相互作用。具有荧光标签的蛋白质中单个氨基酸的特定标记将提供有关该残基的局部微环境的信息。用光叠链接氨基酸标记，然后在TSRI的生物质谱实验室进行shot弹枪蛋白质组学，将鉴定在酵母中表达的人孤儿受体中该残基的配体相互作用。秀丽隐杆线虫中使用的非天然氨基酸的光链接链接将用于表征与衰老有关的途径。与秀丽隐杆线虫的合作将与Salk Institute的Andrew Dillin合作。