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

维持我们生命的大部分过程都是由蛋白质完成的。大多数蛋白质不是单独起作用，而是与其他蛋白质合作起作用。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。其中一些互动只持续不到一秒钟，而另一些则持续数天。我们之前制造了一种叫做SpyCatcher的“细菌超级胶”，它形成了迄今为止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，例如加速新疫苗的创造或建立DNA测序诊断。在这个提案中，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，涵盖蛋白质相互作用的全部范围，从一个在几秒钟内断裂的，到另一个在几分钟内断裂的，一直到原来的永远不会断裂的。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是如何根据机械应力而变化的，就像细胞探索周围环境或肌肉收缩时发生的那样。

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.