The M2 proton channel from Influenza A is an established drug target, with multiple functions during the viral lifecycle. Amino
acid mutations, in the residues lining the channel pore, have rendered M2 resistant to previously licensed inhibitors. Given the
propensity for genetic reassortment of Influenza A and history of pandemics due to emergence of novel human strains, M2 has been subject
of numerous structural characterization efforts. Attempts at rational drug design targeting M2 proton channel have been impeded by the
limited number of experimental techniques having capabilities for elucidating atomic level interactions of the protein-ligand complexes in
the native-like membrane mimetic environment. Solid state Nuclear Magnetic Resonance (ssNMR) is a technique that has all of these
capabilities for structural characterization of membrane protein drug targets in lipid bilayers. Coupling ssNMR with Computer Aided Drug
Discovery (CADD) is the precise approach needed to decrease time and resources required to generate novel therapeutics. Here, we present a
first structural characterization of the full length wild type M2 channel in complex with inhibitor, and of the S31N mutant in the apo and
drug-bound state. Through the use of diverse ssNMR experiments we tested stereoselectivity of drug binding in the channel pore, structural
changes due to mutation, and characterized novel inhibitor interactions. Molecular dynamic simulations were performed by our collaborators
and were in good agreement with our experimental findings. Together these results deepen our understanding of the atomic level
interactions stabilizing wild type inhibitors in the channel pore, and structural changes in the mutant leading to loss of compound
efficacy. Most importantly, specific interaction described herein are essential for successful outcomes from structure based CADD and be
used in future computational efforts. / A Dissertation submitted to the Institute of Molecular Biophysics in partial fulfillment of the
Doctor of Philosophy. / Fall Semester 2015. / December 7, 2015. / Drug Discovery, Influenza A, M2, membrane proteins, NMR, solid state / Includes bibliographical references. / Timothy Cross, Professor Directing Dissertation; Gregory B. Dudley, University Representative;
Michael Blaber, Committee Member; Alan G. Marshall, Committee Member; Thomas C. S. Keller, Committee Member.
| Identifer | oai:union.ndltd.org:fsu.edu/oai:fsu.digital.flvc.org:fsu_360552 |
| Contributors | Wright, Anna Kozlova (authoraut), Cross, Timothy A. (professor directing dissertation), Dudley, Gregory B. (university representative), Blaber, Michael (committee member), Marshall, Alan G. (Alan George) (committee member), Keller, Thomas C. S. (committee member), Florida State University (degree granting institution), College of Arts and Sciences (degree granting college), Program in Molecular Biophysics (degree granting department) |
| Publisher | Florida State University, Florida State University |
| Source Sets | Florida State University |
| Language | English, English |
| Detected Language | English |
| Type | Text, text |
| Format | 1 online resource (117 pages), computer, application/pdf |
| Rights | This Item is protected by copyright and/or related rights. You are free to use this Item in any way that is permitted by the copyright and related rights legislation that applies to your use. For other uses you need to obtain permission from the rights-holder(s). The copyright in theses and dissertations completed at Florida State University is held by the students who author them. |
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