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PhD Dissertation_Gunjan Dixit.pdf (3.27 MB)

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Investigating the Structural Dynamics and Topology of Human KCNQ1 Potassium Ion Channel using Solid-State NMR and EPR Spectroscopy

Dixit, Gunjan
http://orcid.org/0000-0002-0248-6126
http://rave.ohiolink.edu/etdc/view?acc_num=miami1563317786011624

Abstract Details

2019, Doctor of Philosophy, Miami University, Cell, Molecular and Structural Biology (CMSB).
Structural studies of membrane proteins are seminal to understand their role in myriad of biological functions. However, studying membrane proteins is challenging due to difficulties in stable expression and purification. Apart from the limitations associated with biochemistry involved in the study of membrane proteins, there is lack of suitable/robust biophysical techniques which have been a roadblock to the progress of this research field. In this dissertation, I have used electron paramagnetic resonance (EPR) and nuclear magnetic resonance (NMR) spectroscopic techniques to study human KCNQ1 potassium ion channel protein. As presented in chapter 2 of the thesis, I have provided a structural topology model KCNQ1-voltage-sensor domain (Q1-VSD) in POPC/POPG lipid bilayer using EPR spectroscopy. Chapter 3 details lipid bilayer interactions of Q1-VSD using solid-state 31P and 2H NMR spectroscopy. In chapter 4, development and characterization of a membrane mimetic using SMALPs is described to study membrane protein(s). Finally, chapter 5 details the successful expression and purification of human KCNQ1100-370 consisting of helices 1-6 using bacterial system. The protein is further characterized using different biochemical and biophysical techniques. Overall, this dissertation provides a detailed structural aspect of human KCNQ1 ion channel. This work also sheds light on the protein’s interaction with lipid bilayer mimetics.
Gary Lorigan (Advisor)
Carole Dabney-Smith (Committee Chair)
Rick Page (Committee Member)
Dominik Konkolewicz (Committee Member)
Haifei Shi (Committee Member)
115 p.
Biochemistry; Biophysics; Molecular Biology
KCNQ1, Q1-VSD, Long-QT Syndrome, EPR, Solid-State NMR ,SMALPs, Protein Overexpression and Purification, TEM

Recommended Citations

Citations

  • Dixit, G. (2019). Investigating the Structural Dynamics and Topology of Human KCNQ1 Potassium Ion Channel using Solid-State NMR and EPR Spectroscopy [Doctoral dissertation, Miami University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=miami1563317786011624

    APA Style (7th edition)

  • Dixit, Gunjan. Investigating the Structural Dynamics and Topology of Human KCNQ1 Potassium Ion Channel using Solid-State NMR and EPR Spectroscopy . 2019. Miami University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=miami1563317786011624.

    MLA Style (8th edition)

  • Dixit, Gunjan. "Investigating the Structural Dynamics and Topology of Human KCNQ1 Potassium Ion Channel using Solid-State NMR and EPR Spectroscopy ." Doctoral dissertation, Miami University, 2019. http://rave.ohiolink.edu/etdc/view?acc_num=miami1563317786011624

    Chicago Manual of Style (17th edition)

miami1563317786011624
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© 2019, all rights reserved.
This open access ETD is published by Miami University and OhioLINK.