Collaborative Research: MFB: Ultra-Fast Development of Portable Small Molecule Sensor-Actuators

合作研究：MFB：便携式小分子传感器执行器的超快速开发

• 批准号: 2128287
• 负责人: Tim Whitehead
• 金额: $ 54.84万
• 依托单位: University of Colorado at Boulder
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-09-01 至 2024-08-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2128287&HistoricalAwards=false
• 关键词: Collaborative; Research; MFB; Ultra; Fast

In this project, funded by the Molecular Foundation of Biotechnology (MFB) initiative and the Chemical Measurement and Imaging program of the Division of Chemistry, and the Biosensing program of the Division of Chemical, Bioengineering, Environmental, and Transport Systems (CBET), Tim Whitehead of the Department of Chemical and Biological Engineering at the University of Colorado-Boulder, Sean Cutler and Ian Wheeldon at the University of California-Riverside, and Francis Peterson at Medical College of Wisconsin together are developing new ways to control biological systems at will using engineered molecular circuitry. To do this, they will evolve and characterize multiple chemical sensors and derive design rules that enable others to rapidly create new chemically controlled biological responses in any organism of interest. This project seeks to understand how this occurs and define the limits of chemical sensing by specific protein receptors. In doing so, their work will have broad impact on diverse areas of biotechnology, including the precise control of cellular therapies, sensing of illicit drugs or environmental contaminants, and the microbial production of metabolites. This project lies at the interface of chemical biology, organic chemistry, cell and biochemical engineering, and structural biology, and is therefore will provide cross-disciplinary training for the students engaged in this science at all levels. The primary goals of their outreach and education programs are to attract young people to careers in STEM (science, technology, engineering and mathematics) and improve training in chemical biology and industrial biotechnology. The outreach plan involves a multi-pronged effort focused on pre-college and community college engagement, engaging undergraduates through discovery-based labs, and through multiple Research Experiences for Undergraduates programs at all three performance sites.The discovery and exploitation of chemical-induced dimerization (CID) modules was transformative for chemical biology and biotechnology. Chemical-induced dimerization systems have enabled users to construct ligand-controlled systems to modulate cellular and biochemical function, but are constrained by a relatively small number of available protein modules and controlling ligands. This project seeks to develop the foundational knowledge required to understand how CID modules can be reprogrammed to recognize new ligands and to define the limits of chemical space that can be recognized by such systems. To accomplish this, the research team will build and screen computationally designed mutant libraries of PYR1 (PYRABACTIN RESISTANCE 1). PYR1 is a plant hormone receptor that functions through a CID mechanism and possesses a malleable ligand-binding pocket. The project will evolve new PYR1-based sensors controlled by a diverse range of drug-like small molecules. Structural, informatic, and biochemical characterization of these sensors will then be used to develop “rules of recognition” for the PYR1 scaffold and build foundational knowledge required for PYR1-based sensor design. In parallel, a rapid yeast-based system to accelerate PYR1-based sensor development will also be developed. Together, these studies are expected to lead to a detailed understanding of sensor design using the PYR1 system and establish a strong foundation for this enabling and generalizable sensor-design technology.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

在该项目中，由生物技术分子基金会（MFB）倡议和化学部门的化学测量和成像项目以及化学，生物工程，环境和运输系统部门的生物传感项目（CBET）资助，科罗拉多大学博尔德分校化学和生物工程系的Tim Whitehead，加州大学河滨分校的Sean Cutler和Ian Wheeldon，和威斯康辛医学院的弗朗西斯·彼得森（Francis Peterson）正在共同开发利用工程分子电路随意控制生物系统的新方法。为了做到这一点，他们将进化和表征多种化学传感器，并推导出设计规则，使其他人能够在任何感兴趣的生物体中快速创建新的化学控制生物反应。该项目旨在了解这种情况是如何发生的，并定义特定蛋白质受体的化学感应极限。在这样做的过程中，他们的工作将对生物技术的各个领域产生广泛的影响，包括细胞疗法的精确控制、对非法药物或环境污染物的感知以及微生物产生代谢物。本项目是化学生物学、有机化学、细胞与生化工程、结构生物学的交叉学科，将为从事本学科的各级学生提供跨学科的培训。他们的推广和教育计划的主要目标是吸引年轻人从事STEM（科学、技术、工程和数学）领域的职业，并改善化学生物学和工业生物技术方面的培训。外展计划包括多管齐下的努力，重点关注大学预科和社区大学的参与，通过基于发现的实验室吸引本科生，并通过在所有三个演出场地开展的本科生研究体验项目。化学诱导二聚化（CID）模块的发现和开发是化学生物学和生物技术的变革。化学诱导的二聚化系统使用户能够构建配体控制系统来调节细胞和生化功能，但受到相对较少数量的可用蛋白质模块和控制配体的限制。该项目旨在开发所需的基础知识，以了解CID模块如何重新编程以识别新的配体，并定义此类系统可以识别的化学空间的限制。为了实现这一目标，研究小组将构建和筛选PYR1 （PYRABACTIN RESISTANCE 1）的计算设计突变文库。PYR1是一种通过CID机制起作用的植物激素受体，具有可弯曲的配体结合袋。该项目将发展新的基于pyr1的传感器，由各种类似药物的小分子控制。这些传感器的结构、信息和生化表征将用于开发PYR1支架的“识别规则”，并建立基于PYR1的传感器设计所需的基础知识。同时，还将开发一种基于酵母的快速系统，以加速基于pyr1的传感器的开发。总之，这些研究预计将导致使用PYR1系统的传感器设计的详细了解，并为这种启用和通用的传感器设计技术奠定坚实的基础。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

A Closed Form Model for Molecular Ratchet-Type Chemically Induced Dimerization Modules

分子棘轮型化学诱导二聚模块的封闭模型

• DOI: 10.1021/acs.biochem.2c00172
• 发表时间: 2022
• 期刊: Biochemistry
• 影响因子: 2.9
• 作者: Steiner, Paul J.;Swift, Samuel D.;Bedewitz, Matthew;Wheeldon, Ian;Cutler, Sean R.;Nusinow, Dmitri A.;Whitehead, Timothy A.
• 通讯作者: Whitehead, Timothy A.