Liquid disordered–liquid ordered phase coexistence in bicelles containing unsaturated lipids and cholesterol
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Magnetically orienting bicelles are often made by combining the long chain phospholipid 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) with the short chain phospholipid 1,2-dicaproyl-sn-glycero-3-phosphocholine (DCPC) in buffer. These bicelles orient with their bilayer normals perpendicular to the external magnetic field. We have examined the phase behaviour of DMPC/DCPC bicelles and the effects of cholesterol and the unsaturated phospholipid 1,2-dipalmitoleoyl-sn-glycero-3-phosphocholine (DPoPC) as a function of temperature using static solid state (2)H nuclear magnetic resonance spectroscopy. As expected, cholesterol has an ordering effect on the long phospholipid chains and this is reflected in the phase behaviour of the bicelle mixtures. Liquid disordered-liquid ordered, fluid-fluid phase coexistence is observed in DMPC/cholesterol/DCPC bicelles with cholesterol mole fractions of 0.13 and higher. DPoPC/DMPC/cholesterol/DCPC bicelles also exhibit two fluid phase coexistence over a broad range of temperatures and compositions. Bicelles can provide a useful medium in which to study membrane bound peptides and proteins. The orientation parallel to the magnetic field is favourable for studying membrane peptides/proteins because information about the orientation of relevant molecular bonds or internuclear vectors can be obtained directly from the resulting (2)H spectra. Lanthanide ions can be used to flip the bicelles to have their bilayer normals parallel to the external magnetic field. Yb(3+) was used to flip the DPoPC/DMPC/cholesterol/DCPC bicelles while Eu(3+) was found to be ineffective at flipping bicelles containing cholesterol in the present work.
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