A Toolbox for Gating Enzyme Activity using Chemistry and Protein Design

使用化学和蛋白质设计来门控酶活性的工具箱

• 批准号: 1506091
• 负责人: Indraneel Ghosh
• 金额: $ 48万
• 依托单位: University of Arizona
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-07-01 至 2019-06-30
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1506091&HistoricalAwards=false
• 关键词: Toolbox; Gating; Enzyme; Activity; using

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.

有了这个奖项，化学部的生命过程化学计划正在资助亚利桑那大学的Indraneel Ghosh博士开发新方法，用于控制涉及翻译后化学修饰的酶的活性，使用蛋白质工程和设计的小分子的组合。蛋白质的生物活性是由许多翻译后修饰控制的，其中小的化学实体被添加到新合成的蛋白质中。 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。 这种相似性排除了用于直接抑制或激活特定酶功能的特定小分子探针的设计。 已经开发了一些优雅的化学遗传学方法来解决这一需求，但控制同一家族中一种以上酶的活性以开启所需的酶来探测生物学或设计新的有用途径仍然具有挑战性。 该提案旨在建立一个对独特分子开关做出反应的酶工具箱。 这个项目在化学和生物学的接口将提供一个丰富的培训基地，本科生和研究生的多样化集合。 该研究还将加快多学科本科课程的发展，重点是现代实验室研究方法和跨学科座谈会，以讨论当前的一些主题。特别是，参与乙酰化和脱乙酰化的酶将被重新设计，以使它们适合小分子控制。 一个新的和一般的方法来控制特定的酶也将开发使用工程变构开关。 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为目标的多种酶的能力。这些方法有可能更好地破译控制生物学的化学串扰，并有助于重新设计具有许多潜在应用的细胞通路。