Structural Biology of Tyrosine Kinase Regulation

酪氨酸激酶调节的结构生物学

• 批准号: 6675279
• 负责人: DAVID A HORITA
• 金额: $ 14.18万
• 依托单位: WAKE FOREST UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-08-01 至 2005-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6675279
• 关键词: SDS polyacrylamide gel electrophoresis; antiAIDS agent; antiarthritic agent; conformation; data collection methodology /evaluation; drug design /synthesis /production; enzyme activity; enzyme structure; hypoglycemic agents; kinase inhibitor; molecular dynamics; nuclear magnetic resonance spectroscopy; phosphorylation; protein tyrosine kinase; radiotracer; structural biology

DESCRIPTION (provided by applicant): The Src-family of nonreceptor kinases includes eight distinct proteins identified in humans. These proteins play wide-ranging roles in signal transduction in a variety of cell types, and are implicated in numerous diseases including cancer, HIV infection, rheumatoid arthritis, and Type I diabetes (DM1). Inhibition of Hck prevents onset of diabetes in animal models, suggesting this protein as a target for anti- DM1 drug development. The plethora of human tyrosine kinases makes design of high-specificity inhibitors difficult. The long term objective of this project is to develop a comprehensive, detailed understanding of the structural basis of Hck regulation which will be of use in the design of highly specific Hck inhibitors. Such drugs should have broad applicability in the treatment of rheumatoid arthritis and DM1. The specific aims of this pilot project are (1) to establish a bacterial expression system to produce Hck suitable for analysis by solution-state NMR spectroscopy and (2) to initiate NMR studies using these molecules. Specific aim 1 entails stable-isotope labeling of recombinant Hck and in vitro phosphorylation. Specific aim 2 entails assignment of 1H, 13C, and 15N resonance of Hck and collection and analysis of backbone 15N relaxation data. These studies will establish the feasibility of comprehensive analysis of tyrosine kinase structure and dynamics using NMR.

描述（由申请人提供）：非受体激酶的SRC家庭包括在人类中鉴定出的八种不同的蛋白质。这些蛋白质在多种细胞类型的信号转导中起着广泛的作用，与许多疾病有关，包括癌症，HIV感染，类风湿关节炎和I型糖尿病（DM1）。 HCK的抑制可防止动物模型中糖尿病的发作，这表明该蛋白是抗DM1药物开发的靶标。大量的人类酪氨酸激酶使高特异性抑制剂的设计变得困难。该项目的长期目标是对HCK调节的结构基础进行全面，详细的了解，该理解将用于高度特定的HCK抑制剂的设计。这样的药物应在类风湿关节炎和DM1治疗中具有广泛的适用性。 该试验项目的具体目的是（1）建立一个细菌表达系统，以产生适合通过溶液状态NMR光谱分析的HCK，（2）使用这些分子开始NMR研究。特定的目标1需要重组HCK和体外磷酸化的稳定异位标记。具体目标2需要HCK的1H，13C和15N共振的分配以及收集和分析15N放松数据。这些研究将确定使用NMR对酪氨酸激酶结构和动力学进行全面分析的可行性。