Bridge 2: Structural Dynamics of ABC Transporter

多药ATP结合盒（ABC）输出者是普遍存在的ABC转运蛋白， 穿过细胞膜的细胞毒性分子。哺乳动物ABC转运蛋白，如P- 糖蛋白（Pgp）和囊性纤维化跨膜传导调节因子（CFTR）， 专门的出口类，发挥重要的生理作用，并与 疾病研究将继续在确定构象运动的总体目标 通过ABC将ATP能量转化为底物转运的机械功 出口商。该设计利用了稳健的方法和途径，并由 第一阶段的核心，以揭示共同点和差异， 基于特定模式选择的三种原型ABC出口商的构象周期 序列和机械分歧。具体目标旨在界定中间 ATP动力运输所需的状态，绘制过渡途径，并阐明 状态之间相互转换的动力学。一个综合的计算和光谱 该方法利用了五名调查员的专门知识，他们同时参加了 核心和有合作的跟踪记录，以实现特定的目标。我们将使用：用途：a 新的计算方法来映射过渡途径，DEER光谱监测 结构域的运动，快速冷冻淬灭时间解决这些运动和sm-FRET， 检测瞬态中间体并测量停留时间。我们的发现将被整合到 模型，以揭示动态如何控制ABC出口商的功能。

Multidrug ATP binding cassette (ABC) exporters are ubiquitous ABC transporters that extrude cytotoxic molecules across cell membranes. Mammalian ABC transporters, such as P- glycoprotein (Pgp) and cystic fibrosis transmembrane conductance regulator (CFTR), are exclusively of the exporter class, play critical physiological roles and are associated with disease. Research will continue on the overarching goal of defining the conformational motions that transduce the ATP energy to the mechanical work of substrate translocation by ABC exporters. The design leverages robust methods and approaches, tested and refined by the Cores in phase I of the consortium, to reveal commonalities and differences in the conformational cycles of three archetypes ABC exporters selected based on specific pattern of sequence and mechanistic divergences. The specific aims seek to define the intermediate states required for ATP-powered transport, map the transition pathways and elucidate the kinetics of interconversion between states. An integrated computational and spectroscopic approach capitalizes on the expertise of five investigators, who concurrently participate in the Cores and have a track record of collaboration, to achieve the specific aims. We will use : a novel computational approach to map transition pathways, DEER spectroscopy to monitor domain movements, rapid-freeze quench to time resolve these movements and sm-FRET to detect transient intermediates and measure dwell times. Our findings will be integrated into models in order to reveal how dynamics control the function of ABC exporters.