Developing novel methods to watch membrane proteins move

我们研究膜蛋白的结构和功能动力学，特别是许多与医学相关的膜转运系统。越来越多的证据表明，膜蛋白并不是单独作用的，而是作为纳米机器组织起来的，它们通过单个组件的协同作用发挥作用，在时间和空间上都观察到了高精度和特异性。我们正在试图揭开这些蛋白质纳米机械的结构和动力学以及它们的功能背后的原理。我们的实验方法侧重于磁共振技术的使用，特别是电子顺磁共振(EPR)和核磁共振(核磁共振)光谱与分子生物学和生化方法的结合。该项目将研究一种特殊的钠转运体Leut，这是一种来自Aquifex aeolicus的小型氨基酸转运体，它是多巴胺、5-羟色胺、去甲肾上腺素和氨基丁酸的人类神经递质转运体的结构同源物。这些人类转运蛋白与几种疾病状态有关，包括抑郁、焦虑和注意力缺陷多动障碍。几种处方药物针对这些转运蛋白，并与可卡因或安非他明等非法街头药物相互作用。然而，亮氨酸转运蛋白是高度动态的，开发具有医学意义的SLC6抑制剂依赖于了解构象状态的分布。最近的结构研究提出了大规模的构象变化，我们的目标是结合最先进的磁共振技术以及使用新型SMA脂质颗粒纯化和稳定蛋白质的技术来探索该蛋白质家族的功能动力学。

We study the architecture and functional dynamics of membrane proteins, especially many medically relevant membrane transport systems. There is increasing evidence that membrane proteins do not act alone, but that they are organised as nano-machineries which function through the concerted action of individual components with high precision and specificity observed in both time and space. We are seeking to unravel the principles underlying the architecture and dynamics of these protein nanomachineries as well as their function. Our experimental approach focuses on the use of magnetic resonance techniques specifically Electron Paramagnetic Resonance (EPR) and Nuclear Magnetic Resonance (NMR) spectroscopy in combination with molecular biological, and biochemical approaches. This project will study a specific sodium symporter transporter LeuT, a small amino-acid transporter from Aquifex aeolicus, which is a structural homologue of the human neurotransmitter transporters for dopamine, serotonin, norepinephrine and amino butyric acid. These human transporters are implicated in several diseased states including depression, anxiety and attention-deficit hyperactivity disorder. Several prescribed medications target these transporters and illicit street drugs like cocaine or amphetamine interact with them.However, the LeuT transporter is highly dynamic and the development of medically relevant SLC6inhibitors relies on understanding the distribution of conformational states. Recent structural studies have proposed large scale conformational changes and we aim to probe the functional dynamics of this protein family using a combination of state-of-the-art magnetic resonance techniques as well as techniques to purify and stabilise the protein using novel SMA lipid particles.