Protein Manipulation and Engineering with an Electronic Biochemical Approach

疾病往往是蛋白质功能失调的结果。它们可能不会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。单蛋白操作技术将被用于研究动态蛋白质-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。学校将积极招收代表性不足和处境不利的学生，以及当地的印第安人。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。将开发一种单分子操作方法。它将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。该方法应允许检测细微的和短暂的构象波动和转变。此外，本项目将探索配体依赖性酶的波动、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过程中扰动和刺激酶活性来控制酶的功能。通过调节振荡速率和驱动力强度，可以证实被驱动的非线性动态酶系统存在随机共振和随机激活。一旦这种激活被建立，它可以用来触发蛋白质的构象动力学和操纵整体蛋白质的活性和功能。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。

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.