NSF-BSF: Macromolecular crowding in vitro and in cells

NSF-BSF：体外和细胞内的大分子拥挤

• 批准号: 2335137
• 负责人: Gary Pielak
• 金额: $ 75万
• 依托单位: University of North Carolina at Chapel Hill
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-02-01 至 2027-01-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2335137&HistoricalAwards=false
• 关键词: NSF; BSF; Macromolecular; crowding; vitro

Protein-protein interactions regulate all cellular functions, yet they are almost always studied in water rather than in the complex and crowded environment that exists inside cells. The results of this project will fill a key gap that prevents a complete description of metabolism. The goal of moving quantitative biophysics from simple solutions to crowded environments, including inside living cells, is a major challenge with important outcomes. The knowledge gained will add to both the fundamental understanding of biology and inform efforts to produce and stabilize protein-based reagents. The work will also facilitate the training of undergraduate and graduate students in the practice of cutting-edge research. All these efforts are key to building the US bioeconomy.The goal of the project is to gain broadly-applicable knowledge about protein- and protein complex- stability under crowded conditions. A new model combining hard and soft interactions claiming to explain macromolecular crowding effects on the free energy, enthalpy and entropy of protein- and protein complex- stability will be tested on three protein systems using two types or crowding molecules. One system is used to assesses protein stability. The other two systems assess simple dimerization and dimerization with folding. The two types of crowding molecules are industrially important synthetic polymers and a series of de novo designed proteins. This project will use detection system of 19F nuclear magnetic resonance spectroscopy and isothermal titration calorimetry. This collaborative US/Israel project is supported by the US National Science Foundation and the Israeli Binational Science Foundation.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.

蛋白质 - 蛋白质相互作用调节了所有细胞功能，但它们几乎总是在水中而不是在细胞内部存在的复杂且拥挤的环境中进行研究。该项目的结果将填补一个关键空白，以防止对新陈代谢的完整描述。将定量生物物理学从简单解决方案转移到包括内部细胞在内的拥挤环境的目标是重要的挑战。获得的知识将增加对生物学的基本理解，并为生产和稳定基于蛋白质的试剂的努力提供了努力。这项工作还将促进在尖端研究的实践中对本科生和研究生的培训。所有这些努力都是建立美国生物经济学的关键。该项目的目的是在拥挤的条件下获得有关蛋白质和蛋白质复杂稳定性的广泛知识。一个新的模型，结合了声称对大分子的人拥挤对蛋白质和蛋白质复合稳定性的自由能，焓和熵的影响的新模型将使用两种类型或拥挤的分子在三种蛋白质系统上进行测试。一种系统用于评估蛋白质稳定性。其他两个系统通过折叠来评估简单的二聚化和二聚化。两种类型的拥挤分子是工业上重要的合成聚合物和一系列从头设计的蛋白质。该项目将使用19F核磁共振光谱和等温滴定量热法的检测系统。美国国家科学基金会和以色列双原则科学基金会的支持。该奖项反映了NSF的法定任务，并使用该基金会的知识分子优点和更广泛的影响审查标准来反映NSF的法定任务。