ELUCIDATING THE KINETIC MECHANISM OF SCAFFOLD-MEDIATED MAP KINASE SIGNALING

阐明支架介导的 MAP 激酶信号传导的动力学机制

• 批准号: 8169796
• 负责人: WENDELL A LIM
• 金额: $ 0.18万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-09-12 至 2011-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8169796
• 关键词: Behavior; Biochemical; Cells; Computer Retrieval of Information on Scientific Projects Database; Environment; Funding; Grant; Individual; Institution; Kinetics; Life; Maps; Mass Spectrum Analysis; Measures; Mediating; Methods; Pathway interactions; Phosphorylation Site; Phosphotransferases; Play; Proteins; Reaction; Regulation; Research; Research Personnel; Resources; Role; Sampling; Scaffolding Protein; Signal Transduction; Signaling Protein; Source; Specificity; Surface; Thermodynamics; United States National Institutes of Health; base; extracellular; physical model; protein complex; research study; response; scaffold; transmission process

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Living cells interact with and respond to their environment through networks of proteins that transduce extracellular signals into specific biochemical responses inside the cell. Specificity within each pathway is essential for proper regulation of diverse cellular behaviors. Scaffold proteins have been suggested to contribute to specificity by tethering individual signaling proteins together into multi-protein complexes. Emerging evidence suggests that scaffold proteins can also play a more direct role in signal transmission by allosterically promoting specific kinase-kinase reactions and by undergoing conformational changes in response to upstream activators. To construct a physical model for how scaffold proteins promote specific kinase-kinase reactions, we will use a combination of kinetic and biophysical approaches. First, we will measure rate constants for individual kinase-kinase reactions in the presence and absence of scaffolds. Second, we will identify key surfaces and interactions required for scaffold action using crystallographic and biochemical methods. A necessary component of this project is the ability to determine if different protein samples have been phosphorylated, and to identify specific sites of phosphorylation in target proteins. The UCSF mass spectrometry facility will be instrumental in obtaining this information. These experiments will provide kinetic and thermodynamic framework for scaffold-mediated signal transduction, which will provide the basis for understanding how specificity is maintained in cell signaling.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 活细胞通过将细胞外信号转换为细胞内部特定的生化反应的蛋白质网络与其环境相互作用并响应其环境。 每种途径内的特异性对于适当调节各种细胞行为至关重要。 已经建议脚手架蛋白通过将单个信号蛋白链接到多蛋白质络合物中，从而有助于特异性。 新兴证据表明，脚手架蛋白还可以通过变构促进特定的激酶 - 激酶反应以及对上游激活剂的响应构象变化来在信号传输中起更直接的作用。 为了构建一个物理模型，用于脚手架蛋白如何促进特异性激酶 - 激酶反应，我们将使用动力学和生物物理方法的组合。 首先，我们将在存在和不存在脚手架的情况下测量单个激酶反应的速率常数。 其次，我们将使用晶体学和生化方法确定脚手架作用所需的关键表面和相互作用。 该项目的必要组成部分是能够确定是否已经磷酸化了不同的蛋白质样品，并确定靶蛋白中磷酸化的特定位点。 UCSF质谱设施将有助于获取此信息。 这些实验将为支架介导的信号转导提供动力学和热力学框架，这将为理解细胞信号中的特异性维持的基础提供基础。