Membrane proteins contribute about 30% of the proteome and about 60% of the drugs in the market target membrane proteins. But only about 1.5% of structures in Protein Data Bank are membrane proteins which reflect the challenges in membrane protein studies. In this dissertation work M2 full-length protein from Influenza A, a drug target to treat Influenza, is studied in lipid bilayer environment. ssNMR, the best technique to date to study membrane proteins at atomic resolution, has been utilized and intermonomer distance measurement is carried out throughout the study. Differently isotopically labeled protein mixtures were utilized for experiments to obtain unambiguous measurements, even though this method significantly reduces the sensitivity. Influence of amantadine, an antiviral drug, towards distinctive residues at different locations of the channel, such as N-terminus channel pore opening, channel interior and C-terminus of the channel has been characterized. M2 is known to be sensitive to its environment and here the influence of factors such as drugs, pH and membrane constituents such as cholesterol is studied. Distance information obtained for M2 full-length is compared with some of the existing M2 structures of truncated constructs and the importance of studying M2 protein as much as its native-like environment is emphasized. Among multiple drug resistant mutations of M2, S31N mutation contribute almost 100% of drug resistant strains observed in flu infected humans. Therefore, studies involving S31N mutation is an essential part of M2 proton channel studies. Hence, multiple experiments has been performed to obtain structural insights of this oligomeric protein and to compare them with that of M2 wild-type protein, in addition to study of its effect to environmental conditions such as pH. / A Dissertation submitted to the Department of Chemistry and Biochemistry in partial fulfillment of the requirements for the degree of Doctor of Philosophy. / Summer Semester 2015. / July 01, 2015. / Amantadine binding, Cholesterol binding of M2 protein, Influenza A M2 protein, Inter-monomer distance measurements, S31N mutant of M2 protein, Solid-state NMR bspectroscopy / Includes bibliographical references. / Timothy A. Cross, Professor Directing Dissertation; Debra A. Fadool, University Representative; William T. Cooper, Committee Member; Alan G. Marshall, Committee Member.
| Identifer | oai:union.ndltd.org:fsu.edu/oai:fsu.digital.flvc.org:fsu_273635 |
| Contributors | Ekanayake, Ekanayake Vindana (authoraut), Cross, Timothy A. (professor directing dissertation), Fadool, Debra Ann (university representative), Cooper, William T. (committee member), Marshall, Alan G., 1944- (committee member), Florida State University (degree granting institution), College of Arts and Sciences (degree granting college), Department of Chemistry and Biochemistry (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 (199 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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