Establishing a structure-function relationship between biomolecular condensates and protein degradation

建立生物分子缩合物与蛋白质降解之间的结构-功能关系

• 批准号: MR/W01632X/1
• 负责人: Janet Kumita
• 金额: $ 180.55万
• 依托单位: University of Cambridge
• 依托单位国家: 英国
• 项目类别: Fellowship
• 财政年份: 2022
• 资助国家: 英国
• 起止时间: 2022 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=MR%2FW01632X%2F1
• 关键词: Establishing; structure; function; relationship; between

Our bodies contain some 100,000 proteins that enable or regulate essentially every biochemical process on which our lives depend. For these proteins to perform their normal roles, the vast majority must remain in their soluble functional states. To maintain this healthy balance, cells have evolved intricate quality control networks that identify aberrant and damaged components and destroy them. For this to occur, cells need to spatially organise their components to promote these specific reactions and processes. Phase separation, such as when oil is mixed with vinegar, is an important method of compartmentalisation used by the cell to cluster together proteins and other biomolecules for a variety of functions including to: 1) increase enzyme reactions, 2) suspend processes to alleviate cellular stress, or 3) concentrate components together for uptake by the cell's waste-disposal machinery. These droplets contain an array of different proteins and other components and, depending on their biological function, they can be very fluid in nature or they can have a more gel-like composition. In the case of some diseases, further compositional changes from a liquid-like state to irreversible solid-like structures can be harmful to the cell. How does the cell control the phase boundaries within live cells and how do they know when to form droplets in the right place at the right time? To answer these questions, it is necessary to understand the factors that control the droplet composition and characteristics. In this project we propose to design and build novel liquid-droplet forming biomolecules that can: (1) be easily manipulated to introduce site-directed changes that impact on the droplet's physical attributes (i.e. changing the fluidity of the droplet) and (2) specifically recruiting other proteins to the droplets in a controllable-manner to evaluate the impact of these binding partners. We will determine how systematic changes to the novel liquid-droplets affect formation and dissolution of the structures, both in the test-tube and inside cells using complementary experimental techniques. By incorporating a recognition site for autophagosome formation (a key process in autophagy - the cell's waste-disposal mechanism), we will monitor how the changes to droplet structure change the cell's ability to dispose of them. With this structure-function relationship established, we will design artificial phase-separating molecules that can drive the removal of any disease-causing proteins from the cell for use as therapeutics to treat disorders such as Parkinson's disease.

我们的身体含有大约100,000种蛋白质，这些蛋白质基本上可以启动或调节我们生命所依赖的每一个生化过程。为了让这些蛋白质发挥正常的作用，绝大多数蛋白质必须保持其可溶的功能状态。为了保持这种健康的平衡，细胞进化出错综复杂的质量控制网络，识别异常和受损的成分并将其销毁。要做到这一点，细胞需要在空间上组织它们的成分，以促进这些特定的反应和过程。相分离，例如当油与醋混合时，是细胞用来将蛋白质和其他生物分子聚集在一起的一种重要的隔间方法，以实现各种功能，包括：1)增加酶反应，2)暂停过程以减轻细胞压力，或3)将组分集中在一起，供细胞的废物处理机械吸收。这些液滴包含一系列不同的蛋白质和其他成分，根据它们的生物学功能，它们可以是非常流动的，也可以具有更像凝胶的成分。在某些疾病的情况下，从液态到不可逆的固态结构的进一步成分变化可能对细胞有害。细胞如何控制活细胞内的相边界，以及它们如何知道何时在正确的时间在正确的位置形成液滴？要回答这些问题，有必要了解控制液滴组成和特征的因素。在这个项目中，我们建议设计和构建新型的液滴形成生物分子：(1)易于操作，以引入影响液滴物理属性的定点变化(即改变液滴的流动性)；(2)以可控的方式将其他蛋白质特异性地招募到液滴中，以评估这些结合伙伴的影响。我们将使用互补的实验技术，确定对新型液滴的系统性变化如何影响试管和细胞内结构的形成和溶解。通过加入自噬形成的识别位点(自噬的关键过程-细胞的废物处理机制)，我们将监测液滴结构的变化如何改变细胞处理它们的能力。随着这种结构-功能关系的建立，我们将设计人工相分离分子，能够推动任何致病蛋白从细胞中移除，用于治疗帕金森氏症等疾病。

项目成果

Phase separation and molecular ordering of the prion-like domain of the Arabidopsis thermosensory protein EARLY FLOWERING 3.

拟南芥热感应蛋白早期开花3的prion样结构域的相分离和分子排序3。

• DOI: 10.1073/pnas.2304714120
• 发表时间: 2023-07-11
• 期刊: PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA
• 影响因子: 11.1
• 作者: Hutin, Stephanie;Kumita, Janet R.;Strotmann, Vivien I.;Dolata, Anika;Ling, Wai Li;Louafi, Nessim;Popov, Anton;Milhiet, Pierre- Emmanuel;Blackledge, Martin;Nanao, Max H.;Wigge, Philip A.;Stahl, Yvonne;Costa, Luca;Tully, Mark D.;Zubieta, Chloe
• 通讯作者: Zubieta, Chloe

Hypochlorite-Induced Oxidation Promotes Aggregation and Reduces Toxicity of Amyloid Beta 1-42

次氯酸盐诱导的氧化促进聚集并降低淀粉样蛋白 β 1-42 的毒性

• DOI: 10.2139/ssrn.4419865
• 发表时间: 2023
• 作者: Wyatt A

Hypochlorite-induced oxidation promotes aggregation and reduces toxicity of amyloid beta 1-42.

• DOI: 10.1016/j.redox.2023.102736
• 发表时间: 2023-07
• 期刊: REDOX BIOLOGY
• 影响因子: 11.4
• 作者: Manucat-Tan, Noralyn B.;Chowdhury, Ashfaq;Cataldi, Rodrigo;Abdullah, Rafaa Zeineddine;Kumita, Janet R.;Wyatt, Amy R.
• 通讯作者: Wyatt, Amy R.

Probing the effects of N-terminal acetylation on a-synuclein structure, aggregation and cytotoxicity.

探讨 N 末端乙酰化对 α-突触核蛋白结构、聚集和细胞毒性的影响。

• DOI: 10.1016/bs.mie.2022.09.003
• 发表时间: 2023
• 期刊: Methods in enzymology
• 作者: Bell R