This subproject is one of many research subprojects utilizing theresources provided by a Center grant funded by NIH/NCRR. The subproject andinvestigator (PI) may have received primary funding from another NIH source,and thus could be represented in other CRISP entries. The institution listed isfor the Center, which is not necessarily the institution for the investigator.We have been developing 2D-ELDOR (2D electron-electron double resonance) techniques to better capture molecular dynamics in complex fluids, particularly in model and biological membranes. The new 'full Sc- method', which corrects the spectral analysis for the phase distortion effects present in the experiments, enhances the sensitivity of 2D-ELDOR in reporting on molecular dynamics in complex membrane environments. That is, instead of performing spectral fitting in the magnitude mode, our new method enables simultaneous fitting of both the real and imaginary components of the Sc- signal. The full Sc- fitting not only corrects the phase distortions in the experimental data but also more accurately determines instrumental dead times. The phase corrections applied to the Sc- spectrum enable the extraction of the pure absorption-mode spectrum, which is characterized by much better resolution than the magnitude-mode spectrum. In the absorption mode, the variation of homogeneous broadening, which reports on the dynamics of the spin probe, can even be observed by visual inspection. This new method is illustrated with results from model membranes of dipalmitoyl-sn-glycero-phosphatidylcholine (DPPC)-Cholesterol binary mixtures, as well as with results from plasma membrane vesicles of mast cells. In addition to the dynamic parameters, which provide quantitative descriptions for membranes at the molecular level, the high-resolution absorption spectra themselves may be used as a 'fingerprint' to characterize membrane phases and distinguish coexisting components in biomembranes. Thus we find that 2D-ELDOR is greatly improved with the new 'full Sc- method' especially for exploring the complexity of model and biological membranes. A paper describing this work is in press.
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