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Deuterium nuclear magnetic resonance in model membrane systems : an investigation of the interaction of a synthetic, amphiphilic polypeptide with charged lipids

University of British Columbia

The theory of the quadrupole interaction in nuclear magnetic resonance spectroscopy and relaxation measurements is presented in detail, with applications to ²H-NMR studies of order and dynamics in bilayers of deuterated lipids. Investigations of lipid-protein interactions in reconstituted membrane systems and intact biological membranes are reviewed. An experimental program is described which uses a synthetic amphiphilic polypeptide, with known geometry and variable length, to isolate questions about the geometrical interpretation of orientational order in lipid-protein interactions. A report is presented of an investigation of the effects of this polypeptide, Lys₂-Gly-Leu₂₀-Lys₂-Ala-amide, on the mixed bilayer system: Dimyristoylphosphatidylcholine Di-per-deuterio-myristoylphosphatidic Acid. The addition of the peptide was found to have little effect (≤5%) on the first and second moments of the distribution of quadrupole splittings in the liquid crystalline phase. Similarly, the spin-lattice relaxation time constants were affected by ≤10% in the liquid crystalline phase. The time constant for the decay of the quadrupole echo decreased dramatically above the phase transition with the addition of peptide, a phenomenon which is explained in terms of the presence of a new slow motion in the lipid-peptide systems. A simple model of the slow motion induced by the peptide is proposed, in which the lipid molecules undergo a rapid exchange between boundary and bulk sites. An effective correlation time is determined from an average over the rotations on each of these sites. Using this model, estimates are made of the change in the second moment brought about by the onset of the rotations, and. of the number of binding sites on the peptide. These estimates are found to be in agreement with independent measurements of the change in the second moment, and the number of binding sites is within the range predicted by simple considerations of charge balance. The change in the lineshape with the variation of the spacing of the pulses in the quadrupole echo experiment was investigated, and it was determined that the transverse relaxation time constants have a slight orientation dependence. It was also determined that the addition of the peptide has no significant effect on the variation of the lineshape. Some experiments which could answer some of the questions raised by these results are suggested.

Text

2010-05-22

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http://hdl.handle.net/2429/24898

Physics

University of British Columbia

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