Chemical and Biological Switches

化学和生物开关

基本信息

批准号：
155110-2012
负责人：
Woolley, Andrew
金额：
$ 6.27万
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：
2015
资助国家：
加拿大
起止时间：
2015-01-01 至 2016-12-31
项目状态：
已结题

来源：
https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=569417
关键词：
Chemical Biological Switches

项目摘要

Complex living systems, such as a developing multicellular organism, remain a barely charted frontier in our molecular understanding of the natural world. One cannot achieve a molecular understanding of such systems solely by studying their isolated components; to capture their essence one must devise methods to study them intact. Optical methods for manipulating molecular structure and function offer the potential to unravel some of the complexity of living systems. They can be used to target individual molecular species at specific sites with very precise timing inside living animals. The power of this approach has been recognized widely ("Method of the Year" in Nature Methods 2010) but there is still a long way to go to make optical control of molecules in vivo generally practical. We propose to create broadly applicable tools to photo-control protein function in living systems. We will take two complementary approaches: (I) We will design and synthesize derivatives of azobenzene that absorb light at visible wavelengths and undergo large conformational changes. Concurrently we will develop a set of rules for how to attach such molecules to proteins so that the azobenzene isomerization drives a functional change in the protein. (II) We will adapt a naturally light-sensitive protein found in bacteria (photoactive yellow protein) for use as a genetically encoded photoswitch. Using a structure-guided protein engineering approach we will make a circular permutant of photoactive yellow protein that can be used to photo-control a diverse array of target proteins. The fundamental development of these optical tools for manipulating proteins will enhance our ability to understand complex living systems.

复杂的生命系统，例如正在发育的多细胞生物体，在我们对自然世界的分子理解中仍然是一个几乎没有绘制的前沿。仅通过研究这些系统的孤立组成部分，人们无法从分子角度理解这些系统。为了捕捉它们的本质，我们必须设计出方法来完整地研究它们。操纵分子结构和功能的光学方法为揭示生命系统的一些复杂性提供了潜力。它们可用于在活体动物体内以非常精确的时间定位特定位点的单个分子种类。这种方法的力量已得到广泛认可（2010 年《自然方法》中的“年度方法”），但要使体内分子的光学控制普遍实用，还有很长的路要走。我们建议创建广泛适用的工具来光控制生命系统中的蛋白质功能。我们将采取两种互补的方法：（I）我们将设计和合成偶氮苯衍生物，它们吸收可见波长的光并发生较大的构象变化。同时，我们将制定一套如何将此类分子附着到蛋白质上的规则，以便偶氮苯异构化驱动蛋白质的功能变化。（II）我们将采用细菌中发现的天然光敏蛋白（光活性黄色蛋白）作为基因编码的光开关。使用结构引导的蛋白质工程方法，我们将制造光活性黄色蛋白质的圆形排列，可用于光控制多种目标蛋白质。这些用于操纵蛋白质的光学工具的基本发展将增强我们理解复杂生命系统的能力。