Eukaryotic incorporation of photoreactive amino acids

描述（由申请人提供）： 非天然氨基酸将在体内掺入G蛋白偶联受体内的特定位置，以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。用荧光标记物对蛋白质内的单个氨基酸进行特异性标记将提供关于该残基的局部微环境的信息。用光交联氨基酸标记，然后由TSRI的生物质谱实验室进行鸟枪蛋白质组学，将鉴定酵母中表达的人类孤儿受体中该残基的配体相互作用。使用掺入C中的非天然氨基酸的光交联。elegans将被用于表征与衰老有关的途径。与C一起工作。elegans将与索尔克研究所的Andrew Dillin合作。

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.