2 H NMR of oriented phospholipid/cholesterol bilayers containing an amphiphilic peptide

Lipid bilayers form the basis of cell membranes and the phase behaviour of the membrane has been linked to proper cell function. Model membranes composed of relatively simple mixtures of phospholipids and cholesterol can already exhibit complex phase behaviour. Specifically, liquid ordered-liquid disordered fluid phase coexistence occurs in mixtures which contain one saturated long chain phospholipid and one unsaturated long chain phospholipid and cholesterol. This fluid-fluid two phase region persists over a broad range of temperatures and sample compositions and can be observed experimentally in various sample preparations including multilamellar dispersions, bicelles, and multi-lamellae stacked on glass slides. In order to explore the practicality of using oriented samples with different concentrations of the peptide, we investigated the effect of the addition of a synthetic 22 residue amphiphilic peptide on the orientability and phase behaviour of the lipid mixtures, as well as the orientation and dynamics of the peptide itself via ²H NMR. Increasing the peptide concentration promoted the formation of the liquid ordered phase, suggesting a preferential interaction of the peptide with the thicker ordered phase. However, higher peptide content (> 4 mol%) had a significant negative effect on the alignment of bicelles with their bilayer normal perpendicular to the external magnetic field. In the stacked bilayer samples, 6 mol% peptide eliminated the two phase coexistence region altogether and a single liquid ordered phase was observed from 285 to 311 K. Even so, ²H spectra of the peptide itself did not reveal any preference for the peptide to partition into either the liquid disordered or liquid ordered phase and we found two populations of the peptide, one which undergoes rapid axial reorientation about the bilayer normal and a second (powder component) which does not.