Chemical and biological photoswitches

化学和生物光电开关

• 批准号: RGPIN-2017-06663
• 负责人: Woolley, Andrew
• 金额: $ 6.56万
• 依托单位: University of Toronto
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2020
• 资助国家: 加拿大
• 起止时间: 2020-01-01 至 2021-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=712176
• 关键词: Chemical; biological; photoswitches

We aim to develop general ways to control protein function using light. In so doing, we make new photoswitchable molecules and learn how to couple these to proteins to drive functional transitions. We explore how natural photo-proteins can evolve new function and we develop in vitro evolution and selection methods to generate tools for chemical biology. We are taking two general approaches to achieve these goals. The first is a 'bottom-up' approach starting with the chromophore and designing photo-proteins de novo. We propose to modify the chromophore azobenzene, which can be reversibly switched between trans and cis states by light, so that it switches with near-infrared light. A near-IR switchable azobenzene core would enable advances in photopharmacology, biomedical engineering and drug delivery. We aim to find general ways to attach azobenzene-based switches to proteins to control protein folding and thereby function. We will use a combinatorial approach to find azobenzene based cross-linkers and cross-linking sites that permit photo-control of fynomers, which are small antibody alternatives. In parallel, we will test a novel bio-orthogonal labelling approach for targeting intracellular proteins, where thiol-based cross-linking is complicated by the presence of glutathione. Our second approach starts with a naturally-occurring photo-proteins, explores how these work and tries to engineer them as elements for driving change in other proteins. In the process of re-engineering natural photo-proteins, we discovered remarkable light-induced structural transitions including domain swapping and beta-strand slippage. These light-driven phenomena reveal surprising fundamental features of protein dynamics. We aim to test where duplication of a beta strand is generally tolerated in beta-sheets and how often this can lead to new function. Finally, we will explore new phage display technologies to select and evolve photo-proteins.

我们的目标是开发利用光控制蛋白质功能的通用方法。通过这样做，我们制造了新的光可切换分子，并学习如何将它们与蛋白质耦合以驱动功能转变。我们探索天然光蛋白如何进化出新功能，并开发体外进化和选择方法来生成化学生物学工具。 我们正在采取两种一般方法来实现这些目标。第一种是“自下而上”的方法，从发色团开始，从头设计发光蛋白。我们建议对发色团偶氮苯进行修饰，使其可以通过光在反式和顺式状态之间可逆地切换，从而使其在近红外光下进行切换。近红外可切换偶氮苯核心将促进光药理学、生物医学工程和药物输送的进步。我们的目标是找到将基于偶氮苯的开关连接到蛋白质上以控制蛋白质折叠并从而发挥功能的通用方法。我们将使用组合方法来寻找基于偶氮苯的交联剂和交联位点，以允许对 fynomer（小型抗体替代品）进行光控制。与此同时，我们将测试一种针对细胞内蛋白质的新型生物正交标记方法，其中基于硫醇的交联因谷胱甘肽的存在而变得复杂。 我们的第二种方法从天然存在的光蛋白开始，探索它们的工作原理，并尝试将它们设计为驱动其他蛋白质变化的元素。在重新设计天然光蛋白的过程中，我们发现了显着的光诱导结构转变，包括结构域交换和β链滑移。这些光驱动现象揭示了蛋白质动力学令人惊讶的基本特征。我们的目标是测试 β 折叠中 β 链重复的耐受性以及这种重复产生新功能的频率。最后，我们将探索新的噬菌体展示技术来选择和进化光蛋白。