Protein Manipulation and Engineering with an Electronic Biochemical Approach

用电子生化方法进行蛋白质操作和工程

• 批准号: 2317111
• 负责人: Yongki Choi
• 金额: $ 35.7万
• 依托单位: North Dakota State University Fargo
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-15 至 2026-07-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2317111&HistoricalAwards=false
• 关键词: Protein; Manipulation; Engineering; Electronic; Biochemical

Diseases are often the result of proteins that are malfunctioning. They may not be interacting with other molecules as they usually do. They might no longer perform their assigned function in a cell or tissue. Measuring the movements of individual proteins would help determine the exact nature of the problem causing the disease. However, capturing the rapid movements of single proteins is difficult. A single-protein manipulation technique will be employed to study the dynamic protein-protein interactions. A range of research-based, active-learning opportunities will be directed towards K-12 students and undergraduates. Underrepresented and disadvantaged students, as well as local Native Americans, will be actively recruited.The goals of this project are to identify the dynamics of protein-ligand interactions and to attempt to manipulate these interactions relative to specific functions. A single-molecule manipulation approach will be developed. It will combine fluorescence assay methods with a high-speed, high-temporal resolution electronic single-molecule nanocircuit. The approach should allow the detection of subtle and transient conformational fluctuations and transitions. In addition, this project will probe ligand-dependent enzyme fluctuations, conformational transitions, and catalysis, and link them through protein-ligand binding models. Furthermore, this project aims to control enzyme functions by perturbing and stimulating enzyme activities during ligand interactions using a weak, non-thermal oscillating force field. Tuning the oscillation rate and the strength of the driving force may confirm the presence of stochastic resonance and stochastic activation in a driven nonlinear dynamic enzyme system. Once such activation is established, it could be used to trigger protein conformational dynamics and manipulate overall protein activities and functions.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.

疾病往往是蛋白质功能失常的结果。它们可能不像通常那样与其他分子相互作用。它们可能不再在细胞或组织中执行指定的功能。 测量单个蛋白质的运动将有助于确定导致疾病的问题的确切性质。然而，捕捉单个蛋白质的快速运动是困难的。单蛋白质操纵技术将被用来研究动态蛋白质-蛋白质相互作用。一系列以研究为基础的主动学习机会将针对K-12学生和本科生。代表性不足和弱势学生，以及当地的美洲原住民，将积极招募。这个项目的目标是确定蛋白质配体相互作用的动力学，并试图操纵这些相互作用相对于特定的功能。将开发单分子操纵方法。它将联合收割机荧光分析方法与高速、高时间分辨率的电子单分子纳米电路相结合。该方法应允许检测微妙的和短暂的构象波动和转变。此外，该项目将探测配体依赖的酶波动，构象转变和催化作用，并通过蛋白质-配体结合模型将它们联系起来。此外，该项目旨在通过使用弱的非热振荡力场在配体相互作用期间扰动和刺激酶活性来控制酶功能。调节振荡速率和驱动力的强度可以证实在被驱动的非线性动力学酶系统中存在随机共振和随机激活。一旦这种激活被建立，它可以被用来触发蛋白质构象动力学和操纵整体蛋白质的活动和functions.This奖项反映了NSF的法定使命，并已被认为是值得通过使用基金会的智力价值和更广泛的影响审查标准进行评估的支持。