Molecular interaction and high-resolution X-ray structures of the subunits of the osmoregulated ABC-transporter OpuA from B. subtillis

枯草芽孢杆菌渗透调节 ABC 转运蛋白 OpuA 亚基的分子相互作用和高分辨率 X 射线结构

• 批准号: 5402518
• 负责人: Professor Dr. Lutz Schmitt
• 金额: --
• 依托单位: Lehrstuhl für Biochemie I
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 财政年份: 2003
• 资助国家: 德国
• 起止时间: 2002-12-31 至 2006-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/5402518?language=en
• 关键词: Molecular; interaction; resolution; ray; structures

The osmoprotectant uptake system OpuA is one of the three osmoprotective transporters in Bacillus subtilis, which mediate the uptake of the compatible solute glycine betaine, an osmoprotectant widely used by bacteria, plants and even human cells. The transporter complex consists of a lipid-anchored substrate-binding protein (OpuAC), an integral transmembrane protein (OpuAB), and an ATPase subunit (OpuAA) and belongs to the super family of ABC-transporters. In recent years, the osmoprotective transporter complex OpuA has been intensively characterized on a genetic and physiological level. However, no in vitro studies have been performed including structural, biochemical or biophysical studies. Based on our biochemical studies performed so far, we propose to study this transporter complex with respect to its structure and the dynamic aspects of the communication between the subunits within the complex. In vitro and in vivo experiments with the OpuA system will accompany our structural investigations. For the in vitro experiments, crystallography, surface plasmon resonance, and fluorescence techniques will be of major importance. These studies will be aided by in vivo experiments using already available deletion strains of the osmo-regulated transporter systems of B. subtilis. Based on the proposed experiments, we hope to provide a detailed understanding of the structural requirements and communication mechanisms governing the function of the OpuA ABC-transporter of B. subtilis.

植物保护剂摄取系统OpuA是枯草芽孢杆菌中的三种植物保护剂转运蛋白之一，其介导相容性溶质甘氨酸甜菜碱的摄取，所述相容性溶质甘氨酸甜菜碱是细菌、植物甚至人类细胞广泛使用的植物保护剂。转运蛋白复合物由脂质锚定底物结合蛋白（OpuAC）、整合跨膜蛋白（OpuAB）和ATP酶亚基（OpuAA）组成，属于ABC转运蛋白超家族。近年来，在遗传和生理水平上，已经密集地表征了抗氧化剂保护性转运蛋白复合物OpuA。然而，尚未进行体外研究，包括结构、生物化学或生物物理学研究。基于我们的生化研究进行到目前为止，我们建议研究这个转运复合体的结构和动态方面的复杂的亚基之间的通信。OpuA系统的体外和体内实验将伴随我们的结构研究。对于体外实验，晶体学，表面等离子体共振和荧光技术将是非常重要的。这些研究将通过使用B的β-调节转运蛋白系统的现有缺失菌株的体内实验来辅助。枯草杆菌。基于所提出的实验，我们希望提供一个详细的理解的结构要求和通信机制的控制功能的OpuA ABC转运B。枯草杆菌。