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

蛋白质如何与其环境沟通对许多生命过程至关重要，而下一代生物技术的发展需要详细了解这一过程的分子知识。变构通讯是蛋白质将环境信号转化为有用反应（如基因表达）的主要手段。了解变压的详细机理是一个棘手的问题。尽管经过了三十多年的研究，这种生物交流的基本模式在分子水平上仍未得到解决。对变构通讯的透彻理解将有助于开发新的蛋白质设计规则。“定制”变构功能的设计有望彻底改变生物技术，即在开发更有效的生物药物、诊断工具和工业流程方面。这些技术进步将通过实现精确和可预测的生物通信来响应所需的环境线索来实现。除了促进我们对变构沟通的理解外，这项研究还将促进前沿研究领域新科学家的发展，并将包括对STEM中代表性不足群体的培训。这一举措将有助于发展一支多样化和敬业的科学和工程劳动力队伍。本研究的目的是破译潜在的分子机制，通过这种机制，变构信号通过交替变构控制的LacI变体穿过支架。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，将为两个或多个语言构建交流的实验图。一个给定谱系的成员将进行生物物理评估，以破译潜在的分子机制。将利用系统发育和生物物理数据来设计LacI支架的替代变构途径，其中精确的性能指标（即调整的动态范围，配体灵敏度和时间响应性）被授予。这项研究将有助于确定LacI支架中的变构是否可以通过多个残基网络授予，或者变构是否需要单个保守的残基网络。完成后，本研究将通过制定特定变构操作的设计规则，对有关交替变构通信的起源和分子力学的断言进行测试。反过来，该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，特别是在合成生物学中。此外，本研究将大大拓宽我们对变构交际的基本认识。

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.