The structural parameters of fluid phase bilayers composed of phosphatidylcholines with fully saturated, mixed, and branched fatty acid chains, at several temperatures, have been determined by simultaneously analyzing small-angle neutron and X-ray scattering data. Bilayer parameters, such as area per lipid and overall bilayer thickness have been obtained in conjunction with intrabilayer structural parameters (e.g. hydrocarbon region thickness). The results have allowed us to assess the effect of temperature and hydrocarbon chain composition on bilayer structure. For example, we found that for all lipids there is, not surprisingly, an increase in fatty acid chain trans-gauche isomerization with increasing temperature. Moreover, this increase in trans-gauche isomerization scales with fatty acid chain length in mixed chain lipids. However, in the case of lipids with saturated fatty acid chains, trans-gauche isomerization is increasingly tempered by attractive chain-chain van der Waals interactions with increasing chain length. Finally, our results confirm a strong dependence of lipid chain dynamics as a function of double bond position along fatty acid chains.
Quantitative structures were obtained for the fully hydrated fluid phases of dioleoylphosphatidylcholine (DOPC) and dipalmitoylphosphatidylcholine (DPPC) bilayers by simultaneously analyzing x-ray and neutron scattering data. The neutron data for DOPC included two solvent contrasts, 50% and 100% D(2)O. For DPPC, additional contrast data were obtained with deuterated analogs DPPC_d62, DPPC_d13, and DPPC_d9. For the analysis, we developed a model that is based on volume probability distributions and their spatial conservation. The model's design was guided and tested by a DOPC molecular dynamics simulation. The model consistently captures the salient features found in both electron and neutron scattering density profiles. A key result of the analysis is the molecular surface area, A. For DPPC at 50 degrees C A = 63.0 A(2), whereas for DOPC at 30 degrees C A = 67.4 A(2), with estimated uncertainties of 1 A(2). Although A for DPPC agrees with a recently reported value obtained solely from the analysis of x-ray scattering data, A for DOPC is almost 10% smaller. This improved method for determining lipid areas helps to reconcile long-standing differences in the values of lipid areas obtained from stand-alone x-ray and neutron scattering experiments and poses new challenges for molecular dynamics simulations.
The observation of lateral phase separation in lipid bilayers has received considerable attention, especially in connection to lipid raft phenomena in cells. It is widely accepted that rafts play a central role in cellular processes, notably signal transduction. While micrometer-sized domains are observed with some model membrane mixtures, rafts much smaller than 100 nm-beyond the reach of optical microscopy-are now thought to exist, both in vitro and in vivo. We have used small-angle neutron scattering, a probe free technique, to measure the size of nanoscopic membrane domains in unilamellar vesicles with unprecedented accuracy. These experiments were performed using a four-component model system containing fixed proportions of cholesterol and the saturated phospholipid 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), mixed with varying amounts of the unsaturated phospholipids 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) and 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC). We find that liquid domain size increases with the extent of acyl chain unsaturation (DOPC:POPC ratio). Furthermore, we find a direct correlation between domain size and the mismatch in bilayer thickness of the coexisting liquid-ordered and liquid-disordered phases, suggesting a dominant role for line tension in controlling domain size. While this result is expected from line tension theories, we provide the first experimental verification in free-floating bilayers. Importantly, we also find that changes in bilayer thickness, which accompany changes in the degree of lipid chain unsaturation, are entirely confined to the disordered phase. Together, these results suggest how the size of functional domains in homeothermic cells may be regulated through changes in lipid composition.
The structural phases of magnetically alignable lipid mixtures were investigated as a function of temperature and lipid concentration using small-angle neutron scattering (SANS). Two systems were examined: (a) an aqueous mixture of DMPC (dimyristoyl phosphatidylcholine) and DHPC (dihexanoyl phosphatidylcholine) lipids doped with Tm3+ ions resulting in the positive alignment of the system with the applied magnetic field and (b) the above aqueous Tm3+ doped lipid mixture containing a negatively charged lipid, DMPG (dimyristoylphosphatidylglycerol). For both systems, three different scattering patterns were observed corresponding to distinct structural phases at specific temperatures and lipid concentrations. At 45 °C and a lipid concentration of >0.05 g/mL, the high-viscosity liquid crystalline phase was found to be a perforated and possibly undulating lamellar phase consistent with NMR results. Upon dilution (<0.05 g/mL) at the same temperature (45 °C), the perforated lamellar phase transformed into a unilamellar vesicular phase, in which the bilayers may also be perforated. Below about 25 °C, the viscosity decreases considerably and the scattering data suggest that the lamellae present at higher temperatures break up into smaller entities characterized by the bicellar morphology proposed previously for the nondoped system. The structural dimensions of the vesicular and bicellar phases have been determined as a function of lipid concentrations from the SANS data. In the lamellar phase, the influence of Tm3+ ions and DMPG on bilayer structure (e.g., lamellar repeat spacing, bilayer rigidity, and magnetic alignment) were also investigated.
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