SpySwitches: switchable SpyCatcher interactions yielding a modular toolbox for biochemistry and cell biology

SpySwitches：可切换的 SpyCatcher 交互，为生物化学和细胞生物学提供模块化工具箱

• 批准号: BB/T004983/1
• 负责人: Mark Howarth
• 金额: $ 52.63万
• 依托单位: University of Oxford
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2020
• 资助国家: 英国
• 起止时间: 2020 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FT004983%2F1
• 关键词: SpySwitches; switchable; SpyCatcher; interactions; yielding

Most of the processes that keep us alive are carried out by proteins. The majority of proteins do not act on their own but work in partnership with other proteins. Therefore, understanding how and where proteins interact is an important challenge. Some of these interactions only last for a fraction of a second, while others can last for days. We previously generated a "bacterial superglue" called SpyCatcher, which forms one of the strongest protein interactions ever found. SpyCatcher has found many uses, such as for accelerating creation of new vaccines or for building diagnostics to sequence DNA. In this proposal we will use evolution and design to engineer the next generation of SpyCatcher technology. We will generate new versions of SpyCatcher that span the full range of protein interactions, from one that breaks in seconds, another that breaks in minutes, all the way to the original that never breaks. We will also make versions of SpyCatcher that respond to acidic conditions or to light, so that researchers can turn the interaction on as desired. This unprecedented control over protein interactions should have many applications for basic research and biotechnology. In particular we will study how interactions in a cell change according to the mechanical stresses, as occurs when cells explore their surroundings or when a muscle contracts.

维持我们生命的大部分过程都是由蛋白质完成的。大多数蛋白质本身并不起作用，而是与其他蛋白质一起起作用。因此，了解蛋白质如何以及在何处相互作用是一个重要的挑战。其中一些相互作用仅持续几分之一秒，而另一些则可以持续数天。我们之前生产了一种名为SpyCatcher的“细菌强力胶”，它形成了有史以来发现的最强的蛋白质相互作用之一。SpyCatcher已经发现了许多用途，例如用于加速新疫苗的创造或用于构建DNA序列的诊断方法。在本提案中，我们将使用进化和设计来设计下一代SpyCatcher技术。我们将生成新版本的SpyCatcher，它涵盖了蛋白质相互作用的全部范围，从几秒钟内中断的一个，到几分钟内中断的另一个，一直到永远不会中断的原始版本。我们还将制作对酸性条件或光线有反应的SpyCatcher版本，以便研究人员可以根据需要打开相互作用。这种对蛋白质相互作用的前所未有的控制应该在基础研究和生物技术方面有许多应用。特别是，我们将研究细胞中的相互作用如何根据机械应力而变化，就像细胞探索周围环境或肌肉收缩时发生的那样。

项目成果

Structural basis for a conserved neutralization epitope on the receptor-binding domain of SARS-CoV-2.

SARS-COV-2的受体结合结构域上保守的中和表位的结构基础。

• DOI: 10.1038/s41467-023-35949-8
• 发表时间: 2023-01-19
• 期刊: NATURE COMMUNICATIONS
• 影响因子: 16.6
• 作者: Huang, Kuan-Ying A.;Chen, Xiaorui;Mohapatra, Arpita;Nguyen, Hong Thuy Vy;Schimanski, Lisa;Tan, Tiong Kit;Rijal, Pramila;Vester, Susan K. K.;Hills, Rory A. A.;Howarth, Mark;Keeffe, Jennifer R. R.;Cohen, Alexander A. A.;Kakutani, Leesa M. M.;Wu, Yi-Min;Shahed-Al-Mahmud, Md;Chou, Yu-Chi;Bjorkman, Pamela J. J.;Townsend, Alain R. R.;Ma, Che
• 通讯作者: Ma, Che

SpySwitch enables pH- or heat-responsive capture and release for plug-and-display nanoassembly.

• DOI: 10.1038/s41467-022-31193-8
• 发表时间: 2022-06-28
• 期刊: Nature communications
• 影响因子: 16.6

Visible Light-Induced Specific Protein Reaction Delineates Early Stages of Cell Adhesion.

• DOI: 10.1021/jacs.3c07827
• 发表时间: 2023-11-15
• 期刊: JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
• 影响因子: 15
• 作者: Rahikainen, Rolle;Vester, Susan K.;Turkki, Paula;Janosko, Chasity P.;Deiters, Alexander;Hytonen, Vesa P.;Howarth, Mark
• 通讯作者: Howarth, Mark