Date of Award
© 2022 Aye K. Kyaw. This work is freely available courtesy of the author. It may be used under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) license. For all other uses, please contact the copyright holder.
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-Share Alike 4.0 International License.
Bachelor of Arts
Chemistry & Biochemistry
Kathleen P. Howard
Membrane proteins play a range of important roles in biological systems, yet they are underrepresented in the data base of high-resolution structures of all proteins. There is intense interest in developing new methodologies for studying membrane proteins. An essential step to membrane protein method development is devising reliable membrane mimics in which to embed membrane proteins. The goal of this thesis was to develop and apply nanodisc membrane mimics to the study of an influenza A membrane protein called M2. Nanodiscs provide a lipid bilayer environment with access to both sides of the bilayer and are smaller than commonly used liposome model membranes whose size provides challenges for some biophysical methods. This thesis shows how the sample composition of M2 containing nanodiscs was optimized. Dynamic light scattering and size exclusion chromatography was used to characterize M2-nanodiscs. Electrophysiological and budding assays showed that M2 in liposomes were in a functionally relevant conformation. Extensive previous work has been done on studying M2 protein in spherical liposome using site-directed spin label electron paramagnetic resonance (SDSL-EPR). We carried out SDSL-EPR studies of M2-nanodiscs and compared them to published work on M2 in liposomes. Our EPR data is consistent with M2 protein in nanodiscs having a similar conformation, mobility and membrane topology as that seen in previously published M2-liposome work. Furthermore, we probed the ability of nanodiscs to allow for conformational exchange by comparing the impact of drug binding on M2-nanodiscs with M2-liposomes.
Kyaw, Aye K. , '22, "Design and Application of Membrane Nanodiscs for Biophysical Studies of Influenza A Proteins" (2022). Senior Theses, Projects, and Awards. 257.