RUI: Investigating Structure and Dynamic Properties of the Potassium Channel Accessory Protein, KCNE3

膜蛋白在生物系统中具有许多重要的功能，包括离子和分子在细胞膜上的运输。膜蛋白的结构动力学研究是生物物理学和结构生物学的前沿领域之一，对理解膜蛋白的功能至关重要。该研究项目将寻求对钾通道辅助蛋白KCNE3的结构和动力学特性的理解，KCNE3是一种调节电压门控钾通道功能的蛋白质，在心脏、神经和听觉系统中发挥作用。此外，这项工作将促进本科生和高中生的教学和培训。青年科学家将参与该项目的实验和计算方面，因此将接受科学、技术、工程和数学（STEM）研究方面的培训，为该国在这一领域培养世界级的人才。这项研究将增加代表性不足的女性学生，包括第一代大学生在STEM研究中的参与度。本科生和高中生将在国内或国际科学会议上展示他们的研究成果。一个教学水平的现代生物物理学课程将被设计来补充和加强坎贝尔斯维尔大学的教育设施。在该项目下，针对当地高中生和家长的科学研讨会和推广计划将有助于在当地社区传播科学意识，促进对这项科学工作和STEM教育和培训的非正式讨论。基础设施，包括在该项目下建立的研究级光谱仪，将扩大PI机构的本科研究设施，并为肯塔基州中部附近的学院和大学提供通道。KCNE3是KCNE家族的单一跨膜蛋白，可调节电压门控钾通道的功能和运输，包括KCNQ1。了解KCNE3的结构和动力学信息对于了解KCNE3与钾通道蛋白的相互作用及其在通道门控过程中的功能非常重要。尽管KCNE3的重要性显而易见，但关于该系统的信息却很少。本研究的目的是利用电子顺磁共振（EPR）光谱技术和分子动力学建模来研究嵌入在膜中的KCNE3的结构和动力学特性。这项研究填补了目前该领域的空白，并将导致对钾通道的结构和功能的基本理解。双电子-电子共振（DEER） EPR谱将用于测量KCNE3上自旋标签之间的远程距离（25 ~ 80 Å），以观察KCNE3在胶束和脂质双分子层中的结构构象变化。将基于EPR数据建立KCNE3在脂质双分子层中的结构模型。本研究将重点回答以下重要的生物学意义问题：膜双层中KCNE3的结构和拓扑结构是什么？与胶束中的构象相比，KCNE3在膜双分子层中的构象是什么？不同膜环境下KCNE3和KCNE3/KCNQ1的分子运动是如何改变的？本科生和高中生将接受尖端分子生物学科学技术的培训，研究结果将在国家/国际会议上发表，并在同行评审的科学期刊上发表。该项目由分子和细胞生物科学（MCB）部门和促进竞争性研究的既定计划（EPSCoR）共同资助。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。

Membrane proteins are responsible for many important functions in biological systems including the transport of ions and molecules across cellular membranes. Structural dynamics studies of membrane proteins represent one of the final frontiers in biophysics and structural biology and are essential for understanding membrane protein function. This research project will seek an understanding of the structural and dynamical properties of the potassium channel accessory protein KCNE3, a protein that modulates the function of voltage gated potassium channels that play a role in cardiac, nervous and auditory systems. In addition, the work will promote undergraduate and high-school student teaching and training. Young scientists will be involved in both the experimental and computational aspects of the project and hence be trained in science, technology, engineering and mathematics (STEM) research for the preparation of a world class work force in this field within the country. This research will increase the participation of under-represented and women students including first generation college students in STEM research. Undergraduate and high-school students will present their research results at national or international scientific conferences. A teaching-level modern biophysics course will be designed to complement and enhance educational facilities at Campbellsville University. A science workshop and outreach program developed under this project and aimed at local high school students and parents will help spread scientific awareness and promote informal discussions of this scientific work and STEM education and training within the local community. The infrastructure, including a research-level spectrometer established under this project, will expand the undergraduate research facilities at the PI’s institution and also provide access to nearby colleges and universities in Central Kentucky. KCNE3 is a single transmembrane protein of the KCNE family that modulates the function and trafficking of voltage gated potassium channels including KCNQ1. Information on the structure and dynamics of KCNE3 is very important in understanding the interaction of KCNE3 with the potassium channel protein and its function during channel gating. Despite the clear importance of KCNE3, very little information about this system exists. The objective of this research is to investigate structural and dynamical properties of KCNE3 embedded in a membrane utilizing electron paramagnetic resonance (EPR) spectroscopic techniques and molecular dynamics modeling. This research fills a current gap in the field and will lead to a fundamental understanding of the structure and function of the potassium channels. The double electron-electron resonance (DEER) EPR spectroscopy will be used to measure long-range distances (25 to 80 Å) between spin labels located on KCNE3 in order to observe structural conformational changes of KCNE3 in micelle and lipid bilayers. A structural model of KCNE3 in lipid bilayers will be developed based on EPR data. This study will focus on answering the following important biologically significant questions: What is the structure and topology of KCNE3 in membrane bilayers? What is the conformational state of KCNE3 in membrane bilayers when compared to that in micelles? How is the molecular motion of KCNE3 and KCNE3/KCNQ1 altered in different membrane environments? Undergraduate and high-school students will be trained in cutting-edge molecular biological scientific techniques and the results will be presented at national/international conferences and published in peer-reviewed scientific journals.This project is jointly funded by Molecular and Cellular Biosciences (MCB) Division and the Established Program to Stimulate Competitive Research (EPSCoR).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.