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

RUI：研究钾通道辅助蛋白 KCNE3 的结构和动态特性

• 批准号: 2040917
• 负责人: Indra Sahu
• 金额: $ 29.7万
• 依托单位: Campbellsville University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-03-01 至 2025-02-28
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2040917&HistoricalAwards=false
• 关键词: RUI; Investigating; Structure; Dynamic; Properties

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.

膜蛋白在生物系统中负责许多重要功能，包括跨细胞膜的离子和分子的运输。膜蛋白的结构动力学研究代表了生物物理学和结构生物学的最后前沿之一，对于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KCNE 3的结构和动力学特性的理解，KCNE 3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一流的劳动力做好准备。这项研究将增加代表性不足的女学生，包括第一代大学生在STEM研究中的参与。本科生和高中生将在国家或国际科学会议上展示他们的研究成果。将设计一个教学级的现代生物物理学课程，以补充和加强坎贝尔斯维尔大学的教育设施。在该项目下开发的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传播科学意识，并促进当地社区内对这项科学工作和STEM教育和培训的非正式讨论。该基础设施，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，并提供进入肯塔基州中部附近学院和大学的机会。KCNE 3是KCNE家族的一种单跨膜蛋白，调节包括KCNQ 1在内的电压门控钾通道的功能和运输。KCNE 3的结构和动力学信息对于理解KCNE 3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KCNE 3的重要性显而易见，但关于这个系统的信息却很少。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模型研究嵌入在膜中的KCNE 3的结构和动力学性质。这项研究填补了该领域目前的空白，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光谱将用于测量位于KCNE 3上的自旋标记之间的长程距离（25至80 μ m），以观察KCNE 3在胶束和脂质双层中的结构构象变化。基于EPR数据，将建立KCNE 3在脂质双层中的结构模型。本研究将集中于回答以下重要的生物学意义的问题：什么是KCNE 3的结构和拓扑结构的膜双层？与胶束相比，KCNE 3在膜双层中的构象状态是什么？KCNE 3和KCNE 3/KCNQ 1的分子运动在不同的膜环境中是如何改变的？本项目由分子和细胞生物科学部（MCB）和促进竞争性研究既定计划（EPSCoR）共同资助，将对大学生和高中生进行尖端分子生物科学技术的培训，并将在国家/国际会议上发表成果，并在同行评审的科学期刊上发表。该奖项反映了NSF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。

项目成果

Comparing the structural dynamics of the human KCNE3 in reconstituted micelle and lipid bilayered vesicle environments

• DOI: 10.1016/j.bbamem.2022.183974
• 发表时间: 2022-10-01
• 期刊: BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES
• 影响因子: 3.4
• 作者: Campbell, Conner;Faleel, Fathima Dhilhani Mohammed;Sahu, Indra D.
• 通讯作者: Sahu, Indra D.

Electron paramagnetic resonance spectroscopic characterization of the human KCNE3 protein in lipodisq nanoparticles for structural dynamics of membrane proteins

• DOI: 10.1016/j.bpc.2023.107080
• 发表时间: 2023-07-31
• 期刊: BIOPHYSICAL CHEMISTRY
• 影响因子: 3.8
• 作者: Scheyer，Matthew W.;Campbell，Conner;Sahu，Indra D.
• 通讯作者: Sahu，Indra D.

Studying Conformational Properties of Transmembrane Domain of KCNE3 in a Lipid Bilayer Membrane Using Molecular Dynamics Simulations

• DOI: 10.3390/membranes14020045
• 发表时间: 2024-02-01
• 期刊: MEMBRANES
• 影响因子: 4.2
• 作者: Moura，Anna Clara Miranda;Asare，Isaac K.;Sahu，Indra D.
• 通讯作者: Sahu，Indra D.

Perspective on the Effect of Membrane Mimetics on Dynamic Properties of Integral Membrane Proteins

• DOI: 10.1021/acs.jpcb.2c07324
• 发表时间: 2023-04-20
• 期刊: JOURNAL OF PHYSICAL CHEMISTRY B
• 影响因子: 3.3
• 作者: Sahu，Indra D.;Lorigan，Gary A.
• 通讯作者: Lorigan，Gary A.

Role of membrane mimetics on biophysical EPR studies of membrane proteins

• DOI: 10.1016/j.bbamem.2023.184138
• 发表时间: 2023-02-11
• 期刊: BIOCHIMICA ET BIOPHYSICA ACTA-BIOMEMBRANES
• 影响因子: 3.4
• 作者: Sahu，Indra D.;Lorigan，Gary A.
• 通讯作者: Lorigan，Gary A.