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 Ã…) 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.
