Understanding trafficking and membrane localization to re-engineer host and GPCR protein for improved expression

了解运输和膜定位以重新设计宿主和 GPCR 蛋白以改善表达

• 批准号: 1249200
• 负责人: Anne Robinson
• 金额: $ 18.07万
• 依托单位: Tulane University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-06-01 至 2014-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1249200&HistoricalAwards=false
• 关键词: Understanding; trafficking; membrane; localization; re

This NSF award by the Biotechnology, Biochemical and Biomass Engineering program supports work to develop improved expression systems for integral membrane proteins. Integral membrane proteins play critical roles in cell signaling, and are implicated in a wide range of human diseases such as asthma, obesity, cancer, heart disease, and chronic pain. Biophysical and biochemical studies of membrane proteins require large quantities of purified protein; even more than soluble proteins, because they need to be solubilized in detergent solutions. Results for many classes of membrane proteins suggest that in traditional expression systems, valuable targets of drug discovery and biochemical importance will be understudied because researchers cannot make sufficient protein. The goal of this research effort is to improve the expression of an important class of membrane proteins, the G protein-coupled receptor family (GPCRs). Specific aims are to identify cellular and molecular features of both the proteins and expression host to achieve the highest level of active GPCRs for structural and biophysical characterization. In addition, specific educational examples will be developed for the undergraduate Heat and Mass Transfer course. Graduate and undergraduate students will be trained in the laboratory, with particular efforts focused on representation from women and under-represented groups.

这项由生物技术、生物化学和生物质工程项目授予的国家科学基金会奖支持开发改进的整体膜蛋白表达系统的工作。整体膜蛋白在细胞信号传导中起着至关重要的作用，并涉及广泛的人类疾病，如哮喘、肥胖、癌症、心脏病和慢性疼痛。膜蛋白的生物物理和生化研究需要大量的纯化蛋白；甚至比可溶性蛋白质还要多，因为它们需要在洗涤剂溶液中溶解。许多类膜蛋白的结果表明，在传统的表达系统中，由于研究人员无法制造足够的蛋白质，因此对药物发现和生化重要性的有价值靶标的研究将不足。这项研究的目标是提高一类重要膜蛋白的表达，即G蛋白偶联受体家族（gpcr）。具体目的是鉴定蛋白质和表达宿主的细胞和分子特征，以实现最高水平的活性gpcr结构和生物物理表征。此外，将为本科热与质传递课程开发具体的教学实例。研究生和本科生将在实验室接受培训，特别注重妇女和代表性不足群体的代表性。