A BACTERIAL FACTORY FOR PRODUCTION OF MEMBRANE PROTEINS

生产膜蛋白的细菌工厂

基本信息

批准号：
6786562
负责人：
PHILIP D LAIBLE
金额：
$ 25.65万
依托单位：
UNIVERSITY OF CHICAGO
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-08-01 至 2006-07-31
项目状态：
已结题

项目摘要

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.

描述：（逐字研究申请人的摘要）膜和嵌入其中的蛋白质将专用机械划分为提供了细胞和细胞器传达，产生能量的手段，通过运输来吸收营养，排泄废物，转导信号内部隔间之间的代谢物，并建立离子的梯度（和其他小分子），用于为所有正常的细胞活性加油健康的生物。膜蛋白的结构信息非常稀缺 - 众所周知，很难净化数量的本地材料足以进行结晶尝试。今天的方法蛋白质过量生产无法处理膜蛋白，因此重要的类几乎被忽略。为了取得重大进步需要进行创新的策略，而不是增量的进步现有技术。我们利用了杜鹃花的独特生理光合细菌过表达异源蛋白的种类，并最近表明人类外膜蛋白被表达，并且掺入该生物的诱导膜中。我们现在建议开发该系统是功能膜表达的一般系统蛋白质 - 来自任何生物的蛋白质 - 足以生化研究和结晶试验进行结构确定。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Sequences of versatile broad-host-range vectors of the RK2 family.

DOI：
10.1016/s0147-619x(03)00030-1
发表时间：
2003-07
期刊：
Plasmid
影响因子：
2.6
作者：
H. N. Scott;P. Laible;D. Hanson
通讯作者：
H. N. Scott;P. Laible;D. Hanson

Temperature and cryoprotectant influence secondary quinone binding position in bacterial reaction centers.

温度和冷冻保护剂影响细菌反应中心的仲醌结合位置。

DOI：
10.1016/j.febslet.2004.06.042
发表时间：
2004
期刊：
FEBS letters
影响因子：
3.5
作者：
Pokkuluri,PRaj;Laible,PhilipD;Crawford,AdamE;Mayfield,JoyF;Yousef,MohammedA;Ginell,StephanL;Hanson,DeborahK;Schiffer,Marianne
通讯作者：
Schiffer,Marianne

Incorporation of selenomethionine into induced intracytoplasmic membrane proteins of Rhodobacter species.

将硒代蛋氨酸掺入红杆菌属诱导的胞质内膜蛋白中。