| Type of Document |
Master's Thesis |
| Author |
May, Shanna Marie, |
| URN |
etd-07072004-145808 |
| Title |
Gd3+-Nitroxide Pairs for Protein-Membrane Studies:
A Multifrequency EPR Approach
|
| Degree |
Master of Science |
| Graduate Program |
Chemistry |
| Advisory Committee |
| Advisor Name |
Title |
| Dr. Tatyana Smirnova |
Committee Chair |
| Dr. Ed Bowden |
Committee Member |
| Dr. Stefan Franzen |
Committee Member |
|
| Keywords |
- EPR
- distance
- multifrequency
- protein-membrane
- Gd-nitroxide
|
| Date of Defense |
2004-06-30 |
| Availability |
unrestricted |
Abstract
The objective of this research project is to advance the well-established method of Site-Directed Spin Labeling (SDSL) Electron Paramagnetic Resonance (EPR) by utilizing many advantages that High Field EPR (HF EPR) could offer. Specifically, we propose to utilize nitroxide-Gd3+ label pairs for site-directed distance measurements in protein and protein-membrane structure studies. The proposed method is based on through space dipole-dipole interactions between Gd3+ ion positioned on the surface of the bilayer and a nitroxide label attached at the point of interest on the lipid molecule or membrane associated protein. The proposed labeling approach coupled with multifreqency EPR can provide information on Gd3+-nitroxide distance, as well as orientation of Gd3+. Experiments were conducted to investigate the mechanism of nitroxide-Gd3+ interactions in solutions of various viscosities at multiple magnetic fields (corresponding frequencies from 9.5 to 220 GHz) in order to determine relative contributions of dipole-dipole and exchange interactions in nitroxide-Gd3+ pairs at various viscosities and to elucidate the relaxation processes modulating the dipole-dipole interaction as a function of magnetic field. Applications for membrane and membrane protein studies are also discussed.
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| Files |
| Filename |
Size |
Approximate Download Time
(Hours:Minutes:Seconds)
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56K Modem |
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774.61 Kb |
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