STRUCTURE AND DYNAMICS OF EMRE

这个子项目是许多利用资源的研究子项目之一 由NIH/NCRR资助的中心拨款提供。子项目的主要支持 而子项目的主要调查员可能是由其他来源提供的， 包括其它NIH来源。 列出的子项目总成本可能 代表子项目使用的中心基础设施的估计数量， 而不是由NCRR赠款提供给子项目或子项目工作人员的直接资金。 EmrE是一种小型多药耐药转运蛋白，在大肠杆菌中将质子输入耦合到多环芳烃阳离子输出。大肠杆菌，从而赋予对这类药物的抗性。该项目的目标是了解这种二级主动转运蛋白在分子水平上的作用。这需要详细的结构知识，加上动态信息，包括结构变化的时间尺度、幅度和方向。作为一种完整的膜蛋白，这种类型的次级活性转运蛋白是一个挑战。我们以前试图分配骨架酰胺共振阻碍了非常动态，使这个蛋白质有趣。我们现在已经开发了一种新的双胞溶解方案和优化的脂质组合物，以减少动力学和促进分配。这些任务将是至关重要的进一步解释我们的动力学数据和结构确定。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. EmrE is a small multidrug resistance transporter that couples proton import to polyaromatic cation export in E. coli, thus conferring resistance to drugs of this type. The goal of this project is to understand how this secondary active transport protein acts on a molecular level. This requires detailed structural knowledge coupled with dynamic information, including the timescale, amplitude and direction of structural changes. As an integral membrane protein, secondary active transporters of this type are a challenge to work with. Our previous attempts to assign the backbone amide resonances were hampered by the very dynamics that make this protein interesting. We have now developed a new bicelle solubilization protocol and optimized lipid composition to reduce dynamics and facilitate assignment. These assignments will be critical for further interpretation of our dynamics data and structure determination.