A BACTERIAL FACTORY FOR PRODUCTION OF MEMBRANE PROTEINS

描述：（申请人摘要的逐字记录）膜和 它们内部的蛋白质划分了专门的机制， 提供了细胞和细胞器交流，产生能量， 吸收养分，排泄废物，通过运输传递信号 代谢物之间的内部隔室，并建立梯度的离子（和 其他小分子），用于为所有正常的细胞活动提供燃料， 健康的有机体。膜蛋白的结构信息是非常重要的 稀缺-众所周知，要提纯大量的天然物质是非常困难的。 这对于结晶尝试来说是足够的。今天的方法， 蛋白质的过量生产不能处理膜蛋白，因此， 重要的类几乎被忽略了。为了取得重大进展， 需要的是创新战略，而不是渐进式的进步， 现有技术。我们利用红细菌独特的生理学 使光合细菌物种过表达异源蛋白，和 最近已经表明，表达了人外膜蛋白， 被整合到这种生物的诱导膜中。我们现在建议发展 该系统是一个通用的表达功能膜的系统 蛋白质-来自任何生物体-数量足以 用于结构测定的生物化学研究和结晶试验。

Description: (verbatim from the applicant's abstract) Membranes and the proteins embedded within them compartmentalize specialized machinery that provides the means by which cells and organelles communicate, generate energy, take up nutrients, excrete wastes, transduce signals by transporting metabolites between internal compartments, and build gradients of ions (and other small molecules) which are used to fuel all normal cellular activities in healthy organisms. Structural information for membrane proteins is exceedingly scarce - it is notoriously difficult to purify quantities of native material that are sufficient for crystallization attempts. Today's methods for the overproduction of protein cannot deal with membrane proteins, thus this important class is virtually ignored. In order for significant advances to be made, innovative strategies are needed rather than incremental advances in existing technology. We have exploited the unique physiology of the Rhodobacter species of photosynthetic bacteria to overexpress heterologous proteins, and have recently shown that a human outer membrane protein is expressed and incorporated into induced membranes of this organism. We now propose to develop this system to be a general one for the expression of functional membrane proteins - from any organism - in quantities that are sufficient for biochemical studies and crystallization trials for structure determination.