The suprastructure-function relationship between amyloid assemblies and their toxic and infectious potentials

淀粉样蛋白组装体及其毒性和传染性潜力之间的超结构-功能关系

• 批准号: BB/S003312/1
• 负责人: Wei-Feng Xue
• 金额: $ 46.29万
• 依托单位: University of Kent
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2018
• 资助国家: 英国
• 起止时间: 2018 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FS003312%2F1
• 关键词: suprastructure; function; relationship; between; amyloid

A number of human disorders, for example Alzheimer's disease (AD), Parkinson's disease (PD), and transmissible spongiform encephalopathies (TSEs), are associated with the abnormal folding and assembly of proteins. The net result of this misfolding is the formation of large insoluble protein deposits as well as toxic and transmissible protein particles in a state called amyloid. Not all amyloids are associated with disease, as some are tolerated by the cells or even perform beneficial functions for their host organisms. Why some amyloid are disease-associated and toxic while others are not is a fundamentally important biological question that we currently do have answer to. This gap in our knowledge not only prevents researchers from fully understanding the fundamental biology in the amyloid life-cycle, but also prevent pharmaceutical industries from targeting the correct molecular structures and developing effective therapeutics against the devastating amyloid associated diseases. In this project, we will address this knowledge gap by investigating the new idea that whether amyloid is associated with pathology or not is linked to how individual amyloid filaments, as building blocks, are organised to form large structures in the range of a millionth to a billionth of metre in size, which we call the amyloid suprastructure.Recently, detailed atomic-resolution structural models for several disease-associated amyloid fibrils resolved using solid-state nuclear magnetic resonance spectroscopy and transmission electron microscopy methods have allowed insight into how individual atoms are organised in the amyloid structures. Using these methodologies have resolved the detailed organisation of individual protein chains in amyloid structures, yet how these structures interact with biology and why some amyloid are associated with disease even though all amyloid share the same type of organisation of the protein chains has not been understood. Here, we propose that the "missing-link" between our knowledge on amyloid structures and how they interact with biology and/or associated with disease is encoded in the type of suprastructure individual amyloid filaments will assembly into, that is whether these amyloid building-blocks may form straight bundles, twisted ropes or tubes, large open networks or tightly packed clusters etc. If we can gather data on the types of suprastructures amyloid building-blocks can form and follow how each of these structures may influence how amyloid interact with cells and propagate in a disease context, then we will be able to resolve the missing-link between amyloid structure and their biology. This is exactly what we can now do, as we will use atomic force microscopy (AFM) imaging method that enable us to visualise large number of individual amyloid superstructures that are between millionths of metre in size to billionths of metre in size, so called mesoscopic size range. Atomic force microscopy imaging of amyloid structures in these intermediate length scales between the sizes of atoms to the size of cells gives low-noise and high resolution images that are very much ideal for the precise quantitative measurement of individual amyloid suprastructures in sample that are composed of mixtures of diverse assemblies such as amyloid samples. This combined with biological measurements of how the same amyloid structures behave with/in cells, we will be uniquely placed to discover the missing-link between structure of amyloid and their cellular functions. Our findings will shed new light on the why amyloid structures can confers cytotoxicity and infectivity in mammals and humans in some but not all cases, and will also give us clues as to what pharmaceutical industries should target in the search for effective therapies against the devastating diseases some amyloid structures are associated with.

许多人类疾病，例如阿尔茨海默病（AD）、帕金森病（PD）和传染性海绵状脑病（TSE），都与蛋白质折叠和组装异常有关。这种错误折叠的最终结果是形成大量不溶性蛋白质沉积物以及处于称为淀粉样蛋白状态的有毒且可传播的蛋白质颗粒。并非所有淀粉样蛋白都与疾病相关，因为有些淀粉样蛋白可以被细胞耐受，甚至对其宿主生物体发挥有益的功能。为什么有些淀粉样蛋白与疾病相关且有毒，而另一些淀粉样蛋白则不然，这是一个根本性的重要生物学问题，我们目前确实有答案。我们知识上的这一差距不仅阻碍了研究人员充分了解淀粉样蛋白生命周期的基础生物学，而且还阻碍了制药行业瞄准正确的分子结构并开发针对破坏性淀粉样蛋白相关疾病的有效疗法。在这个项目中，我们将通过研究新的想法来解决这一知识差距，即淀粉样蛋白是否与病理学相关，与作为构建块的单个淀粉样蛋白丝如何组织形成百万分之一到十亿分之一米大小的大型结构（我们称之为淀粉样蛋白上层结构）有关。最近，针对几种与疾病相关的淀粉样蛋白原纤维的详细原子分辨率结构模型 使用固态核磁共振波谱和透射电子显微镜方法解决这一问题，使人们能够深入了解单个原子在淀粉样蛋白结构中的组织方式。使用这些方法已经解决了淀粉样蛋白结构中各个蛋白链的详细组织问题，但这些结构如何与生物学相互作用以及为什么某些淀粉样蛋白与疾病相关，尽管所有淀粉样蛋白都具有相同类型的蛋白链组织，但尚不清楚。在这里，我们提出，我们对淀粉样蛋白结构的了解以及它们如何与生物学相互作用和/或与疾病相关的知识之间的“缺失环节”被编码在单个淀粉样蛋白丝将组装成的上层结构类型中，即这些淀粉样蛋白构件是否可以形成直束、绞绳或管、大型开放网络或紧密堆积的簇等。如果我们能够收集有关上层结构类型的数据 淀粉样蛋白构件可以形成并遵循这些结构如何影响淀粉样蛋白与细胞相互作用以及在疾病背景下传播，然后我们将能够解决淀粉样蛋白结构与其生物学之间缺失的联系。这正是我们现在可以做的，因为我们将使用原子力显微镜（AFM）成像方法，使我们能够可视化大量个体淀粉样蛋白超结构，其尺寸在百万分之一米到十亿分之一米之间，即所谓的介观尺寸范围。对介于原子大小和细胞大小之间的中等长度尺度的淀粉样蛋白结构进行原子力显微镜成像，可提供低噪声和高分辨率的图像，非常适合精确定量测量由不同组件（例如淀粉样蛋白样品）的混合物组成的样品中的单个淀粉样蛋白超结构。结合相同淀粉样蛋白结构在细胞内/细胞内行为的生物学测量，我们将处于独特的地位，可以发现淀粉样蛋白结构与其细胞功能之间缺失的联系。我们的研究结果将揭示为什么淀粉样蛋白结构在某些但不是所有情况下可以在哺乳动物和人类中赋予细胞毒性和感染性，并且还将为我们提供线索，了解制药行业在寻找针对与某些淀粉样蛋白结构相关的破坏性疾病的有效疗法时应瞄准哪些目标。

项目成果

Amyloid particles facilitate surface-catalyzed cross-seeding by acting as promiscuous nanoparticles.

• DOI: 10.1073/pnas.2104148118
• 发表时间: 2021-09-07
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: Koloteva-Levine N;Aubrey LD;Marchante R;Purton TJ;Hiscock JR;Tuite MF;Xue WF
• 通讯作者: Xue WF

Structural Identification of Individual Helical Amyloid Filaments by Integration of Cryo-Electron Microscopy-Derived Maps in Comparative Morphometric Atomic Force Microscopy Image Analysis.

• DOI: 10.1016/j.jmb.2022.167466
• 发表时间: 2022-04-15
• 期刊: JOURNAL OF MOLECULAR BIOLOGY
• 影响因子: 5.6
• 作者: Lutter, Liisa;Al-Hilaly, Youssra K.;Serpell, Christopher J.;Tuite, Mick F.;Wischik, Claude M.;Serpell, Louise C.;Xue, Wei-Feng
• 通讯作者: Xue, Wei-Feng

Structural reconstruction of individual filaments in Aß42 fibril populations assembled in vitro reveal rare species that resemble ex vivo amyloid polymorphs from human brains

对体外组装的 A42 原纤维群中单个纤维的结构重建揭示了类似于人脑离体淀粉样蛋白多态性的稀有物种

• DOI: 10.1101/2023.07.14.549001
• 发表时间: 2023
• 作者: Aubrey L

Quantification of amyloid fibril polymorphism by nano-morphometry reveals the individuality of filament assembly

通过纳米形态测量法对淀粉样蛋白原纤维多态性进行定量揭示了丝组装的个体性

• DOI: 10.1101/2020.01.14.905877
• 发表时间: 2020
• 作者: Aubrey L

Quantification of amyloid fibril polymorphism by nano-morphometry reveals the individuality of filament assembly.

• DOI: 10.1038/s42004-020-00372-3
• 发表时间: 2020-09-11
• 期刊: COMMUNICATIONS CHEMISTRY
• 影响因子: 5.9
• 作者: Aubrey, Liam D.;Blakeman, Ben J. F.;Lutter, Liisa;Serpell, Christopher J.;Tuite, Mick F.;Serpell, Louise C.;Xue, Wei-Feng
• 通讯作者: Xue, Wei-Feng