EAGER: Examining the Origins and Molecular Pathways of Alternate Allosteric Networks in the Lacl System

EAGER：检查 Lacl 系统中替代变构网络的起源和分子途径

• 批准号: 1747439
• 负责人: Corey Wilson
• 金额: $ 30万
• 依托单位: Georgia Tech Research Corporation
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-08-01 至 2019-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1747439&HistoricalAwards=false
• 关键词: EAGER; Examining; Origins; Molecular; Pathways

How proteins communicate with their environment is critical to many life processes and detailed molecular knowledge of this process is required for the development of next-generation biotechnologies. Allosteric communication is the principal means by which proteins translate an environmental signal into a useful response (e.g., gene expression). Understanding the detailed mechanism of allostery is a vexing problem. Despite more than three decades of study, this fundamental mode of biological communication remains unsolved at the molecular level. A thorough understanding of allosteric communication will facilitate the development of new protein design rules. The design of "custom made" allosteric functions promises to revolutionize biotechnology, namely with regard to the development of more competent biological drugs, diagnostic tools, and industrial processes. These technological advances will be achieved by enabling precise and predictable biological communication in response to desired environmental cues. In addition to advancing our understanding of allosteric communication, this study will enable the development of new scientists in a cutting-edge research area, and will include the training of underrepresented groups in STEM. This initiative will contribute to the development of a diverse and engaged science and engineering workforce.The goal of this study is to decipher the underlying molecular mechanism by which allosteric signals traverse the scaffold across LacI variants with alternate allosteric control. This study will be accomplished by the construction and characterization of a synthetic phylogenetic tree of alternate allosteric networks. To complement inferred evolutionary relationships, experimental maps of communication will be constructed for two or more linages. Members of a given linage will be evaluated biophysically to decipher the underlying molecular mechanism. Phylogenetic and biophysical data will be leveraged to design alternate allosteric routes in the LacI scaffold in which precise performance metrics (i.e., tuned dynamic range, ligand sensitivity and temporal responsiveness) are conferred. This study will help identify whether allostery in the LacI scaffold can be conferred via multiple networks of residues, or whether allostery requires a single conserved network of residues. Upon completion, this study will enable the testing of assertions with regards to the origin and molecular mechanics of alternate allosteric communication via the development of design rules for specific allosteric operations. In turn, this algorithm will produce novel LacI-based transcription factors for use in a broad range of biotechnological applications, specifically in synthetic biology. In addition, this study will significantly broaden our fundamental understanding of allosteric communication.

蛋白质如何与其环境进行通信对于许多生命过程至关重要，并且需要详细的分子知识来开发下一代生物技术。变构通讯是蛋白质将环境信号转化为有用反应的主要手段（例如，基因表达）。了解变构的详细机制是一个令人烦恼的问题。尽管经过了三十多年的研究，这种生物通讯的基本模式在分子水平上仍然没有得到解决。对变构通讯的深入理解将有助于新的蛋白质设计规则的发展。“定制”变构功能的设计有望使生物技术发生革命性变化，即关于更有能力的生物药物、诊断工具和工业过程的开发。这些技术进步将通过响应所需的环境线索实现精确和可预测的生物通信来实现。除了促进我们对变构通信的理解外，这项研究还将促进尖端研究领域的新科学家的发展，并将包括对STEM中代表性不足的群体的培训。这一举措将有助于发展一个多元化的和从事科学和工程workings.The研究的目标是破译的潜在的分子机制，变构信号穿越支架跨LacI变体与替代变构控制。本研究将通过构建和表征交替变构网络的合成系统发育树来完成。为了补充推断的进化关系，将为两个或多个谱系构建通信的实验地图。将对给定谱系的成员进行生物药理学评价，以解读潜在的分子机制。系统发生和生物物理学数据将被用来设计LacI支架中的替代变构途径，其中精确的性能指标（即，调节的动态范围、配体灵敏度和时间响应性）。这项研究将有助于确定是否可以通过多个残基网络赋予LacI支架中的变构，或者变构是否需要一个单一的保守残基网络。完成后，这项研究将能够通过开发特定变构操作的设计规则来测试关于替代变构通信的起源和分子机制的断言。 反过来，这种算法将产生新的基于LacI的转录因子，用于广泛的生物技术应用，特别是合成生物学。此外，这项研究将大大拓宽我们对变构通讯的基本理解。