State-of-the-art ESR for biological applications

在所有的生命形式中，细胞都被一层含有蛋白质和脂类的膜所包围。细胞所需的所有信息，从营养输入、废物输出到环境感知，都需要通过这个外膜。了解通过称为受体的蛋白质进行的交流是一项主要的研究活动，主要是因为所有的信号分子(如激素、药物、抗生素)都需要作用于明确定义的受体。如果我们要在治疗中针对这些受体，制药公司和生物化学家需要了解这些受体在正常和疾病状态下是如何发挥作用的。此外，这些受体蛋白对单个分子的激活很敏感，但可以在很短的时间内(在百万分之一秒内)将这种相互作用放大许多倍(S到数百万倍)--因此，这些蛋白质也有潜力在技术上(生物纳米技术)用于记忆存储、放大、光检测和过滤/筛选等设备中。所有这些工作的问题是缺乏结构信息来允许我们描述受体的功能。我们没有这些受体如何工作的完整图片的原因有很多，但事实证明非常成功的一种获取信息的方法是使用探针的位点定向(SPIN)标记，它可以提供关于距离和受体激活后结构变化的时间尺度的非常精确的信息。需要特殊的新方法和精密的(电子自旋共振)设备，使用最复杂和现代的电子和计算机程序来捕获和分析数据。我们研究的大脑受体可以从大肠杆菌中制造和提纯，负责释放激素和刺激剂来控制食欲，以及肌肉活动。它在控制肥胖、阿尔茨海默氏症和帕金森氏症方面发挥着关键作用--因此它是一个重要的药物靶点，但目前还没有药物可用。在过去的10年里，我们一直在描述这种特殊的受体，并对其了解很多。我们现在已经准备好将我们对该受体的专业知识与新的结构方法结合起来，这些新的结构方法已经在世界各地的几个实验室的2到3个其他膜蛋白上进行了精炼，使用了新的可用的检测设备。这次访问美国是为了让申请者(AW)了解如何做到这一点-AW在该地区非常活跃了19年(1971-1990)，然后进入了相关的(核磁共振)技术。只有通过在开创性的实验室工作，与学生和研究人员交谈，开发想法，一个人才能精通一种新的方法。当AW回来时，牛津大学新资助的设备(最先进的)现在可以用于这项工作。牛津大学的两名学生正在为该项目制造蛋白质(包括探针的突变体)，他们将从引进的专业知识中受益，新成立的(自2007年春季以来)牛津ESR社区的其余10个小组也将受益，其中3个小组将需要新方法。这种协作和交流访问在国际科学中对于保持尖端思想的进步至关重要，这是可以通过在这里请求的支持来实现的。

In all life-forms, cells are bounded by a membrane which contains proteins and lipids. All the information that is required by the cell, from nutrients input, waste product output to environmental sensing, needs to pass through this outer membrane. Understanding communication through proteins called receptors is a major research activity, mainly because all signalling molecules (such as hormones, drugs, antibiotics) need to act on well-defined receptors. Drug companies and biochemists need to understand how these receptors function in both normal and diseased states if we are to target them in therapy. Additionally, these receptor proteins are sensitive to activation by a single molecule but can amplify this interaction many (1000's to millions) times over in very short timeframe (in a millionth of a second) - for this reason, these proteins could also have potential in technology (bionanotechnology) for use in devices for memory storage, amplification, photodetection and filtering/screening, to name a few examples.The problem in all this work is the lack of structural information to permit us to describe how the receptors function. There are many reasons why we do not have such complete pictures of how these receptors work, but one way to get this information, which is proving very successful, is called site-directed (spin) labelling using probes which can give very precise information about distance and the time-scales of structural changes once a receptor is activated. Special new methods and sophisticated (electron spin resonance) equipment is required, with the data being captured and analysed using the most sophisticated and modern electronics and computer programmes.The brain receptor we study can be made and purified from E. coli bacteria, and is responsible for release of hormones and stimulants for appetite control, as well as muscle activity. It plays a pivotal role in the control of obesity, Alzheimer's and Parkinson's diseases - it is therefore a vital drug target but no drugs are yet available. Over the last 10 years, we have been characterising this particular receptor and know a lot about it. We are now ready to combine our expertise of this receptor together with the new structural methods which have been refined on 2 or 3 other membrane proteins in a handful of laboratories worldwide, using newly available detection equipment. This visit to the US is to bring the applicant (AW) back up to date with how to do this - AW was very active in the area for 19 years (1971-1990) and then moved into a related (NMR) technique. Only through working in a pioneering laboratory, talking to students and researchers and developing ideas, can someone become proficient at a new method. Newly funded equipment (state of the art) is now available for this work in Oxford when AW returns. Two students in Oxford are making the protein for the project (mutants to include the probes) and will benefit from the imported expertise, as will the rest of the newly established (since Spring 2007) Oxford ESR community of 10 groups, 3 of which will have need of the new methods. Such collaborative and exchange visits are vital in international science to keep cutting edge ideas progressing, and this is something which can be accomplished using the support requested here.