Probing natural amyloid fibril assembly by protein display

通过蛋白质展示探测天然淀粉样原纤维组装

• 批准号: BB/D019109/1
• 负责人: Paul Barker
• 金额: $ 41.76万
• 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: 2007
• 资助国家: 英国
• 起止时间: 2007 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FD019109%2F1
• 关键词: Probing; natural; amyloid; fibril; assembly

The structure and substance of living things are made from self-assembling molecules, primarily proteins. Protein aggregates have become notorious for being associated with some diseases of old age such as Alzheimer's and Parkinson's Diseases. In these disorders, proteins mis-fold and aggregate into large rope-like deposits known as amyloid fibrils. It turns out that most if not all proteins can fold to form these amyloid filaments. The fibrils have a very regular structure and some remarkable physical properties. It turns out that Nature has evolved some proteins to be in this state deliberately, to make fibrils on bacterial cell surfaces, to help bacteria stick to other cells. These are some of the most stable fibrils of their type. The regular and stable structure of amyloid fibrils makes them a good scaffold on which to hang other, functional proteins, and that is this basis of this proposal. We will use the proteins that form the natural amyloid fibrils on bacterial cells and fuse them, at the level of their genes, to other proteins and enzymes. To do this, we need to understand more about how individual proteins, which are about 2 nanometres in all dimensions, assemble to form fibrils that are 10 nanometres wide and 10 micrometres long. We can alter the protein's sequence and observe how the fibrils change. Once we have established more about how the building blocks are assembled, we will fuse other proteins to the building blocks so that their function becomes displayed on the fibrils. To start with we will display proteins that carry electrons around in cells. This way we can make a conducting amyloid fibril. We will also display on the fibrils enzymes that destroy antibiotics. This can be used to protect the cell against an antibiotic, giving the cell an advantage. This property can be used to evolve the fibrils to be more useful technologically. Once we have established the best type of fibril and protein display methods, we can then make fibrils at will and use them to build up networks of fibrils on the nanometre scale. This is a new approach to building molecular devices for electronics and diagnostics, out of completely self-assembling molecules. So while amyloid is a serious problem in many disease states, it may also be remarkably useful for making molecular circuits.

生物的结构和实质是由自组装分子（主要是蛋白质）制成的。蛋白质骨料因与某些老年疾病（例如阿尔茨海默氏症和帕金森氏病）相关而臭名昭著。在这些疾病中，蛋白质折叠和聚集成大的绳状沉积物，称为淀粉样蛋白原纤维。事实证明，大多数蛋白质都可以折叠以形成这些淀粉样细丝。原纤维具有非常规的结构和一些显着的物理特性。事实证明，大自然已经进化了一些蛋白质，故意在这种状态下，在细菌细胞表面上制作原纤维，以帮助细菌粘在其他细胞上。这些是其类型中最稳定的原纤维。淀粉样蛋白原纤维的常规结构使它们成为悬挂其他功能性蛋白质的良好脚手架，这就是该建议的基础。我们将使用在细菌细胞上形成天然淀粉样蛋白原纤维的蛋白质，并将其在其基因水平上融合到其他蛋白质和酶。为此，我们需要更多地了解各个维度上约2纳米蛋白的单个蛋白质如何组装成10纳米宽和10微米长的原纤维。我们可以改变蛋白质的序列，并观察原纤维的变化。一旦我们建立了更多关于如何组装构建块的方法，我们将融合其他蛋白质与构建块，以便它们的功能显示在原纤维上。首先，我们将显示蛋白质，这些蛋白质在细胞中携带电子。这样，我们可以制作导电淀粉样蛋白纤维。我们还将显示在破坏抗生素的原纤维酶上。这可用于保护细胞免受抗生素的影响，从而使细胞具有优势。该特性可用于发展原纤维以在技术上更有用。一旦我们建立了最佳类型的原纤维和蛋白质显示方法，就可以随意制作原纤维，并使用它们在纳米尺度上建立原纤维网络。这是一种通过完全自组装分子来构建电子和诊断分子设备的新方法。因此，尽管淀粉样蛋白在许多疾病状态下是一个严重的问题，但它对于制作分子电路也可能非常有用。