REGULATION OF SIGNALING BY SCAFFOLD PROTEINS

支架蛋白对信号传导的调节

• 批准号: 6518824
• 负责人: LISAN L PARKER
• 金额: $ 2.28万
• 依托单位: VANDERBILT UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-03-01 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/6518824
• 关键词: Drosophilidae; SDS polyacrylamide gel electrophoresis; affinity chromatography; autoradiography; biological signal transduction; central nervous system; confocal scanning microscopy; electrophysiology; immunofluorescence technique; immunoprecipitation; laboratory rat; lysophospholipase; nerve /myelin protein; nuclear matrix; protein kinase C; protein protein interaction; protein structure function; serotonin; serotonin receptor; transcription factor; yeast two hybrid system

The long term objective of this proposal is to understand how signal transduction pathways operate, specifically, the regulation of signal transduction by scaffold proteins. The specific aims of this research proposal are to: (I). examine the distribution of MUPP1 (multi-PDZ domain protein 1), in the central nervous system by indirect immunofluorescence. (II) investigate the interaction and localization of MUPP1, with components involved in the serotonin 5-HT2C receptor-mediated pathway in the rat brain; (III) evaluate if MUPP1 PDZ domains of highest homology to INAD interact with the three INAD-interacting proteins from Drosophila. This work hopes to contribute to the growing consensus that cells are not merely cytoplasmic bags of molecules that randomly collide and allow various intracellular events to occur. On the contrary, cells tightly organize and regulate various protein:protein interaction events which lead to efficient signal transduction processes. This view of highly regulated cellular organization will increase the number of targets available for drugs to intervene and disrupt the processes of carcinogenesis, neurodegenerative diseases, as well as countless other human afflictions. Indirect immunofluorescence using the confocal microscopy will be employed (aim I). Biochemical techniques including immunoprecipitation, ligand overlay assays, and affinity chromatography will be utilized to investigate protein:protein interaction (aim II, III. Disruption of the mapped sites of interaction on MUPP1 by mutagenesis and reintroduction of these modified constructs of MUPP1 into a 5-HT2C expressing cell line will be used to evaluate the functional consequence the interaction.

该提案的长期目标是了解信号转导途径如何运作，具体来说，通过脚手架蛋白对信号转导的调节。 该研究建议的具体目的是：（i）。通过间接免疫荧光检查中枢神经系统中MUPP1（多PDZ结构蛋白1）的分布。 （ii）研究MUPP1的相互作用和定位，与大鼠脑中5-羟色胺5-HT2C受体介导的途径有关的成分； （iii）评估最高同源性的MUPP1 PDZ域是否与果蝇的三种InaD Interactacting蛋白相互作用。 这项工作希望有助于日益增长的共识，即细胞不仅是随机碰撞并允许发生各种细胞内事件的分子的细胞质袋。 相反，细胞紧密组织和调节各种蛋白质：蛋白质相互作用事件，从而导致有效的信号转导过程。 高度调节的细胞组织的这种观点将增加可用于干预和破坏癌变过程，神经退行性疾病以及无数其他人类苦难的过程的靶标数量。 使用共聚焦显微镜的间接免疫荧光将采用（AIM I）。 生化技术将利用包括免疫沉淀，配体覆盖测定和亲和力色谱法来研究蛋白质：蛋白质相互作用（AIM II，III。通过诱变对MUPP1相互作用的相互作用的破坏，通过诱变作用和重新构造的MUPP1构造将MUPP1的构造造成了5-HT2的互动构建，以将其用于评估型细胞的作用。