Solution NMR spectroscopy studies of an adrenergic receptor b1AR

肾上腺素受体 b1AR 的溶液核磁共振波谱研究

• 批准号: BB/K01983X/1
• 负责人: Daniel Nietlispach
• 金额: $ 57.78万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2013
• 资助国家: 英国
• 起止时间: 2013 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FK01983X%2F1
• 关键词: Solution; NMR; spectroscopy; studies; adrenergic

Living organisms are made up of a very large quantity of cells. Each of these cells contains machinery that is essential to maintain and develop the life of a particular organism. These cells are surrounded by a waterproof lipid membrane, which encapsulates the mostly aqueous interior of a cell that includes also the essential molecular machinery. Every process of life both on a large as well as on a small scale involves continuous adaptations to a changing environment. Following such changes and responding to the demands that arise through the activities of the organisms the conditions within the individual cells need to be continuously adjusted. Every cell needs to be supplemented with nutrients for energy and building materials, waste products need to be removed and instructions need to be given for the multitude of different processes to act in a concerted manner. To facilitate these requirements across the impenetrable lipid membrane a large number of proteins are embedded into the cell membrane. These proteins connect the cell exterior with the inside of the cell and are called membrane proteins. A particular group of these proteins is responsible for relaying information in form of control signals across the membrane. The cells are using these proteins as sensors that relay a message from the exterior to the inside of a cell, where a cell is then able to understand what adjustments need to be made. The range of such control signals can be very diverse and there are therefore several hundreds of these sensors making this a particularly important group of membrane proteins. In fact it turns out to be the largest family of proteins in humans. Our work is looking in more details at these proteins, the so-called G protein coupled receptors GPCRs. We are trying to understand how exactly it is that these proteins work and in particular how different external control signals for a given sensor facilitate the different responses on the inside of the cell. Exposure of these proteins on the surface of the cells makes them easily accessible, which is crucial for them to work properly. It makes them also ideal targets for drugs in situations when our body malfunctions and needs drug therapeutic help. Therefore next to the academic interest in understanding how these proteins work there is a large interest from the pharmaceutical industry for the development of newer and better drugs from which our general well being will benefit.To be able to address such questions typically requires biologists and chemists to zoom in on a molecular level using a range of biophysical techniques, which allow us to see what is happening on an atomic scale. Our lab is using a technique called nuclear magnetic resonance (NMR), which allows us to study these GPCR proteins in a nearly native environment. For the technique to work the GPCR under study is removed from the cell membrane but is still kept surrounded by a very small portion of it. These proteins are extremely unstable and hence very tricky to study. We are concentrating on a particular member of the GPCR family, which has been modified and displays enhanced properties to assist our investigations.These sensor proteins are considered to be highly mobile and their dynamic nature strongly influences how they function. NMR is an excellent method that can describe which parts of these proteins are flexible. We are particularly interested in studying how the mobility in these proteins changes in the presence of different external signals so that we can correlate these variations with the given responses inside the cell. Most likely our results will allow us to make conclusions that are very general in nature as it is highly likely that other GPCRs will function following a similar way of action. So far GPCRs have been elusive to such studies and our work intends to generate this novel insight.

有生命的有机体是由大量的细胞组成的。这些细胞中的每一个都含有维持和发展特定有机体生命所必需的机械。这些细胞被防水的类脂膜包裹着，它包裹着细胞内部的大部分是水的，其中也包括必要的分子机制。生命的每一个过程，无论是大的还是小的，都需要不断地适应不断变化的环境。在这样的变化之后，并对生物体活动产生的需求作出反应，单个细胞内的条件需要不断地调整。每个电池都需要补充能量和建筑材料的营养，需要清除废物，需要指示多种不同的过程以协调一致的方式行动。为了促进这些要求穿过不可穿透的类脂膜，大量的蛋白质被嵌入细胞膜。这些蛋白质连接细胞外部和细胞内部，称为膜蛋白。这些蛋白质中的一组负责通过膜以控制信号的形式传递信息。这些细胞使用这些蛋白质作为传感器，将信息从细胞外部传递到细胞内部，然后细胞能够了解需要进行哪些调整。这种控制信号的范围可能非常不同，因此有数百个这样的传感器，使其成为一组特别重要的膜蛋白。事实上，它被证明是人类最大的蛋白质家族。我们的工作是更详细地研究这些蛋白质，即所谓的G蛋白偶联受体GPCRs。我们正在试图了解这些蛋白质到底是如何工作的，特别是特定传感器的不同外部控制信号如何促进细胞内部的不同反应。这些蛋白质暴露在细胞表面使它们很容易接触到，这对它们正常工作至关重要。当我们的身体出现故障并需要药物治疗帮助时，它也使它们成为药物的理想靶子。因此，除了对了解这些蛋白质如何工作的学术兴趣之外，制药业对开发更新更好的药物也有很大的兴趣，我们的整体福祉将从中受益。为了能够解决这些问题，通常需要生物学家和化学家使用一系列生物物理技术放大分子水平，使我们能够看到原子尺度上正在发生的事情。我们的实验室正在使用一种名为核磁共振(核磁共振)的技术，它使我们能够在几乎自然的环境中研究这些GPCR蛋白。为了使这项技术发挥作用，正在研究的gpr被从细胞膜上移除，但仍然被细胞膜的一小部分包围着。这些蛋白质极不稳定，因此很难研究。我们专注于GPCR家族中的一个特定成员，它已经被修改并显示出增强的性质来帮助我们的研究。这些传感器蛋白被认为具有高度的移动性，它们的动态性质强烈地影响它们的功能。核磁共振是一种很好的方法，可以描述这些蛋白质的哪些部分是灵活的。我们特别感兴趣的是研究这些蛋白质在不同外部信号存在时的流动性如何变化，以便我们能够将这些变化与细胞内给定的反应相关联。最有可能的是，我们的结果将使我们能够得出非常笼统的结论，因为其他GPCR很可能将遵循类似的行动方式发挥作用。到目前为止，GPCR对于这样的研究是难以捉摸的，我们的工作旨在产生这种新的见解。

项目成果

Application of random coherence order selection in gradient-enhanced multidimensional NMR

随机相干阶次选择在梯度增强多维核磁共振中的应用

• DOI: 10.1088/1742-6596/699/1/012004
• 发表时间: 2016
• 期刊: Conference Series
• 作者: Bostock M

Integral membrane protein structure determination using pseudocontact shifts.

使用伪接触位移测定整体膜蛋白结构。

• DOI: 10.1007/s10858-015-9899-6
• 发表时间: 2015
• 期刊: Journal of biomolecular NMR
• 影响因子: 2.7
• 作者: Crick DJ

Fast NMR Data Acquisition - Beyond the Fourier Transform

快速 NMR 数据采集 - 超越傅里叶变换

• DOI: 10.1039/9781782628361-00267
• 发表时间: 2017
• 作者: Bostock M

An Adaptable Phospholipid Membrane Mimetic System for Solution NMR Studies of Membrane Proteins.

• DOI: 10.1021/jacs.7b06730
• 发表时间: 2017-10-25
• 期刊: Journal of the American Chemical Society
• 影响因子: 15
• 作者: Chien CH;Helfinger LR;Bostock MJ;Solt A;Tan YL;Nietlispach D
• 通讯作者: Nietlispach D

Compressed sensing: Reconstruction of non-uniformly sampled multidimensional NMR data

• DOI: 10.1002/cmr.a.21438
• 发表时间: 2017-03-01
• 期刊: CONCEPTS IN MAGNETIC RESONANCE PART A
• 影响因子: 0.6
• 作者: Bostock, Mark;Nietlispach, Daniel
• 通讯作者: Nietlispach, Daniel