New Chemical Tools for Optically Controlled Protein Modification

项目摘要/摘要 光促进的生物相容化学转化已成为化学研究中的有力工具 生物学通过实现对活动的时空控制。虽然基因编码的光激活 工具已经成为生物正交工具箱中的中流砥柱，光驱动共轭方法有效地 在生物相关条件下与天然生物分子结构(无遗传编码)的界面，是 相对有限。在这个项目中，我们将发展一种由我们的团队最近为 氧化还原活性N-氨基甲酰基吡啶盐与色氨酸残基的光生物结合 光致电子转移中的色氨酸。我们将展示这一点，通过仔细调制光学和电化学 这些试剂的性质，我们将能够(1)极大地增强这一动力学能力 标记反应和(2)使得能够发现促进这种标记的新的机制范式 化学反应。此外，我们将通过光学和电化学的仔细操作来证明 N-氨基甲酰基吡啶盐试剂的性质，我们将能够调用对色氨酸的机械控制 以依赖于波长的方式进行标记(即，我们可以控制具有给定波长的反应机理 光)。反过来，这将允许我们设计新的基于应用的实验，既可以精确地 反应结果，显著扩展了光生物偶联化学的能力。具体来说，我们将 利用这种光学和机械控制来设计新的基于活动的传感应用以及 通过设计我们应用于研究知之甚少的过程的邻近标记方法 在线粒体动力学中。

Project Summary/Abstract Biocompatible chemical transformations that are promoted by light have become powerful tools in chemical biology by virtue of enabling spatiotemporal control over activity. Whilst genetically encoded photoactivatable tools have become mainstays in the bio-orthogonal toolbox, light driven conjugation methods that effectively interface with native biomolecular structures (no genetic encoding) under biologically relevant conditions, are comparatively limited. In this project, we will evolve a method recently developed by our group for the photobioconjugation of Tryptophan (Trp) residues using redox-active N-carbamoylpyridinium salts that engage Trp in photo-induced electron transfer. We will show that, by carefully modulating the optical and electrochemical properties of these reagents, that we will be able to both (1) dramatically enhance the kinetic capabilities of this labelling reaction and (2) enable the discovery of new mechanistic paradigms that promote this labelling chemistry. Moreover, we will demonstrate that, through careful manipulation of optical and electrochemical properties of the N-carbamoyl pyridinum salt reagent, that we will be able to invoke mechanistic control over Trp labelling in a wavelength-dependent fashion (i.e. we can control reaction mechanism with a given wavelength of light). This, in turn, will allow us to design new application-based experiments that can both command precise reaction outcomes and markedly expand the capabilities of photobioconjugation chemistry. Specifically, we will harness this optical and mechanistic control for the design of new activity-based sensing applications as well as through the design of proximity labelling approaches that we apply to the study of poorly understood processes in mitochondrial dynamics.