Chemical and Biological Switches

化学和生物开关

• 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

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.

复杂的生活系统，例如发展中的多细胞生物，仍然是我们对自然世界的分子理解中几乎没有图表的前沿。一个人不能仅通过研究其孤立的组件来实现对此类系统的分子理解。为了捕捉他们的本质，必须设计方法来完整研究它们。操纵分子结构和功能的光学方法为揭示了生活系统的某些复杂性提供了潜力。它们可用于靶向具有非常精确的活物时机的特定位点的单个分子物种。这种方法的力量已被广泛认可（《自然方法》 2010年的“年度方法”），但是要在体内对分子进行光学控制通常可以实用，还有很长的路要走。 我们建议在生活系统中创建广泛适用的工具来进行照片控制蛋白功能。我们将采用两种互补方法：（i）我们将设计和合成偶氮苯的衍生物，这些衍生物在可见的波长下吸收光并经历巨大的构象变化。同时，我们将制定一套有关如何将这种分子连接到蛋白质上的规则，以使偶氮苯同构化驱动蛋白质的功能变化。 （ii）我们将适应细菌（光活性黄色蛋白）中的自然光敏蛋白，以用作遗传编码的照片开关。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的圆形置换物，可用于拍摄各种靶蛋白。这些操纵蛋白质的光学工具的基本开发将增强我们了解复杂生活系统的能力。