Protein Manipulation and Engineering with an Electronic Biochemical Approach

疾病往往是蛋白质功能失调的结果。它们可能不会像往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其指定的功能。测量单个蛋白质的运动将有助于确定导致这种疾病的问题的确切性质。然而，捕捉单一蛋白质的快速运动是困难的。将使用单蛋白质操纵技术来研究蛋白质-蛋白质之间的动态相互作用。一系列以研究为基础的主动学习机会将面向K-12学生和本科生。这个项目的目标是确定蛋白质-配体相互作用的动态，并试图操纵这些与特定功能相关的相互作用。将开发一种单分子操作方法。它将结合荧光分析方法和高速、高时间分辨率的电子单分子纳米电路。该方法应允许检测微妙和瞬时的构象波动和转变。此外，该项目将探索依赖于配体的酶的波动、构象转变和催化，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、非热的振荡力场来扰动和刺激配体相互作用中的酶活性来控制酶的功能。调节振荡速率和驱动力的强度可以证实驱动的非线性动态酶系统中存在随机共振和随机激活。一旦这种激活被确立，它就可以用来触发蛋白质构象动力学，并操纵整个蛋白质的活动和功能。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势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.