A BACTERIAL FACTORY FOR PRODUCTION OF MEMBRANE PROTEINS

描述：(逐字摘自申请人的摘要)膜和 嵌入其中的蛋白质将特殊的机械隔开， 提供了细胞和细胞器交流、产生能量、 吸收营养，排泄废物，通过运输传递信号 内部隔室之间的代谢物，并建立离子的梯度(和 其他小分子)，它们被用来为所有正常的细胞活动提供燃料 健康的生物。膜蛋白的结构信息非常丰富。 稀缺--众所周知，要提纯大量的原生材料是非常困难的 对于结晶尝试是足够的。今天的方法是为了 过量生产的蛋白质不能处理膜蛋白，因此 重要的类实际上被忽略了。为了取得重大进展， 需要制定创新的战略，而不是在 现有技术。我们利用了红细菌独特的生理学。 过量表达异源蛋白的光合细菌种类，以及 最近发现了一种人类外膜蛋白的表达和 并入这种生物的诱导膜中。我们现在提议开发一种 该体系有望成为功能膜表达的通用体系 蛋白质-来自任何生物体-的数量足以 用于结构确定的生化研究和结晶试验。

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.