Fish Electric Organ as a Factory for Membrane Proteins

描述(申请人提供)：结构生物学的一个目标是使蛋白质结构测定像现在的DNA测序一样成为常规。通过主要由蛋白质结晶学家和核磁共振光谱仪积累的丰富经验，这一目标对可溶性蛋白质来说是触手可及的。人膜蛋白实现这一目标的主要障碍是目前可用的异源表达系统无法为原子级结构方法学常规生产足够数量的正确折叠的人膜蛋白。人类膜蛋白原子级结构的稀缺是一个问题，因为膜蛋白经常是旨在改善人类健康的药物的靶标。这一提议的假设是，产生大量内源性膜蛋白的真核细胞是生产大量正确折叠的人膜蛋白的新型异源蛋白表达系统的良好候选者。这项提议将用来自鱼类电子器官的细胞来验证这一假设，这些细胞是烟碱型乙酰胆碱受体的异常高水平的生产者。与公共卫生相关：缺乏关于人类膜蛋白形状的原子水平知识是一个问题，因为膜蛋白对人类健康和许多疾病至关重要。膜蛋白允许细胞与外部世界以及细胞内膜分隔的隔室之间进行通信。了解膜蛋白的形状不仅对了解正常细胞的工作方式很重要。这一认识还有助于开发疾病治疗和预防方法，例如，通过指导药物设计。涉及膜蛋白的疾病是主要的公共卫生问题，包括神经退行性疾病、精神疾病和心脏病。

DESCRIPTION (provided by applicant): A goal of structural biology is to make protein structure determination as routine as DNA sequencing is now. That goal is coming within reach for soluble proteins through extensive experience accumulated primarily by protein crystallographers and NMR spectroscopists. The major obstacle to progress toward that goal for human membrane proteins is the inability of currently available heterologous expression systems to routinely produce sufficient amounts of correctly folded, human membrane proteins for atomic level structure methodologies. The scarcity of atomic level structures of human membrane proteins is a problem because membrane proteins are frequent targets for drugs designed to improve human health. The hypothesis for this proposal is that eukaryotic cells producing large amounts of endogenous membrane proteins are good candidates as the foundation for novel heterologous protein expression systems producing large amounts of correctly folded human membrane proteins. This proposal will test this hypothesis with electrocytes from fish electric organ, which are exceptionally high level producers of nicotinic acetylcholine receptors. PUBLIC HEALTH RELEVANCE: The scarcity of atomic level knowledge about the shapes of human membrane proteins is a problem because membrane proteins are crucial to human health and to many diseases. Membrane proteins allow cells to communicate with the external world and between membrane-divided compartments within a cell. Knowing the shapes of membrane proteins is important not only for understanding how normal cells work. This understanding also helps develop disease therapies and preventions, for example, by guiding drug design. Diseases involving membrane proteins are major public health problems including neurodegenerative diseases, mental illnesses, and heart disease.