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.

膜蛋白负责生物系统中的许多重要功能，包括离子和分子跨细胞膜的运输。膜蛋白的结构动力学研究是生物物理学和结构生物学的最终前沿之一，对于理解膜蛋白功能至关重要。该研究项目将寻求了解钾通道附件蛋白KCNE3的结构和动态特性，该蛋白质调节电压门控钾通道的功能，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和高中生教学和培训。年轻的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）的培训，以准备在该国境内该领域的世界一流劳动力。这项研究将增加代表性不足的女学生的参与，包括第一代大学生参与STEM研究。本科生和高中生将在国家或国际科学会议上介绍他们的研究结果。教学级现代生物物理学课程将旨在在坎贝尔斯维尔大学完成和增强教育设施。科学研讨会和宣传计划是根据该项目制定的，针对当地的高中生和父母，将有助于传播科学意识，并促进对当地社区内这种科学工作以及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，并为中部肯塔基州中部的近大学和大学提供通道。 KCNE3是KCNE家族的单个跨膜蛋白，可调节包括KCNQ1在内的电压门控钾通道的功能和运输。有关KCNE3的结构和动力学的信息对于理解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显然很重要，但有关该系统的信息很少。这项研究的目的是研究利用电子顺磁共振（EPR）光谱技术和分子动力学建模的膜中嵌入KCNE3的结构和动态特性。这项研究填补了该领域的当前空白，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光谱法将用于测量位于KCNE3上的自旋标签之间的长距离距离（25至80Å），以观察Micelle和脂质双层中KCNE3的结构变化。脂质双层中KCNE3的结构模型将根据EPR数据开发。这项研究将着重于回答以下重要的生物学意义上的重要问题：膜双层中KCNE3的结构和拓扑是什么？与胶束相比，膜双层中KCNE3的构象状态是什么？在不同的膜环境中，KCNE3和KCNE3/KCNQ1的分子运动如何改变？本科生和高中生将接受尖端分子生物学科学技术的培训，结果将在国家/国际会议上介绍，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家（MCB）的分子（MCB）分层和既定的研究（MCB）奖励的研究（MCB）共同资助，以刺激竞争性研究（Epscore）。诚实地通过评估来诚实地使用基金会的智力优点和更广泛的影响审查标准。

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.

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.

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.