New Chemical Tools for Optically Controlled Protein Modification

项目概要/摘要 由光促进的生物相容性化学转化已经成为化学转化中的有力工具。 生物学通过对活动的时空控制。虽然基因编码的光活化 工具已经成为生物正交工具箱中的支柱，光驱动缀合方法有效地 在生物学相关条件下与天然生物分子结构（无遗传编码）的界面， 相对有限。在这个项目中，我们将发展我们小组最近开发的一种方法， 使用具有氧化还原活性的N-氨基甲酰基吡啶鎓盐的色氨酸（Trp）残基的光生物缀合 光诱导电子转移中的色氨酸。我们将证明，通过仔细调节光学和电化学， 这些试剂的性质，我们将能够（1）显着提高这一动力学能力， 标记反应和（2）能够发现新的机制范式，促进这种标记 化学.此外，我们将证明，通过仔细操纵光学和电化学， 性质的N-氨基甲酰基吡啶盐试剂，我们将能够调用机制控制色氨酸 以波长依赖的方式标记（即，我们可以用给定的波长控制反应机制 光）。反过来，这将使我们能够设计新的基于应用的实验，既可以命令精确， 反应结果，并显着扩大了光生物共轭化学的能力。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， 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.