A Toolbox for Gating Enzyme Activity using Chemistry and Protein Design

通过这一奖项，化学部的生命过程化学计划资助亚利桑那大学的Indrane Ghosh博士开发新的方法，利用蛋白质工程和设计的小分子的组合来控制翻译后化学修饰中涉及的酶的活性。蛋白质的生物活性是由许多翻译后修饰控制的，即在新合成的蛋白质中添加小的化学实体。几乎所有负责催化翻译后修饰的酶都属于具有相似结构的大家族。这种相似性排除了为直接抑制或激活特定酶的功能而设计特定小分子探针的可能性。一些优雅的化学遗传学方法已经被开发出来，以满足这一需求，但为了打开所需的酶来探索生物学或设计新的有用的途径，控制同一家族中多种酶的活性仍然具有挑战性。这项提议寻求建立一个能对独特的分子开关做出反应的酶工具箱。这个位于化学和生物交界处的项目将为不同的本科生和研究生提供丰富的培训场所。这项研究还将加快多学科本科课程的发展，重点放在现代实验室研究方法和讨论一些当前主题的跨学科座谈会上。特别是，参与乙酰化和去乙酰化的酶将被重新设计，使它们易于受小分子控制。还将开发一种新的通用方法来控制特定的酶，使用工程变构开关。从所提出的方法中获得的新知识有可能为社区提供以前所未有的正交控制针对多种酶的能力。这些方法有可能更好地破译支配生物学的化学串扰，并帮助重新设计具有许多潜在应用的细胞途径。

With this award, the Chemistry of Life Processes Program in the Chemistry Division is funding Dr. Indraneel Ghosh from the University of Arizona to develop new methods for controlling the activities of enzymes implicated in posttranslational chemical modifications using a combination of protein engineering and designed small molecules. The biological activity of proteins is controlled by numerous posttranslational modifications, whereby small chemical entities are added to newly synthesized proteins. Almost all enzymes that are responsible for catalyzing posttranslational modifications belong to large families with similar structures. This similarity precludes the design of specific small molecule probes for the direct inhibition or activation of the function of a specific enzyme. A few elegant chemical genetic approaches have been developed to address this need but it remains challenging to control the activity of more than one enzyme within the same family in order to turn-on a desired enzyme to probe biology or engineer new and useful pathways. This proposal seeks to establish a toolbox of enzymes that respond to unique molecular switches. This project at the interface of chemistry and biology will offer a rich training ground for a diverse collection of undergraduate and graduate students. The research will also accelerate the development of multidisciplinary undergraduate courses, with a focus on modern laboratory research methods and cross-disciplinary colloquiums for discussing a number of current topics. In particular, enzymes involved in acetylation and deacetylation will be redesigned for rendering them amenable to small molecule control. A new and general method for controlling specific enzymes will also be developed using an engineered allosteric switch. New knowledge obtained from the proposed approaches has the potential to provide the community with the ability to target multiple enzymes with unprecedented orthogonal control. These methods have the potential to better decipher the chemical cross-talk that governs biology and aid in the reengineering of cellular pathways with many potential applications.