Ternary lipid bilayers containing cholesterol in a high curvature silica xerogel environment.
The phase behavior of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC)/1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC) (1/1 mol ratio)/cholesterol (0-60 mol %) supported lipid bilayers agreed with a DOPC/DSPC/cholesterol ternary phase diagram by Zhao et al. when a mica support was used (Zhao, J.; Wu, J.; Heberle, F. A.; Mills, T. T.; Klawitter, P.; Huang, G.; Costanza, G.; Feigenson, G. W. Biochim. Biophys. Acta, Biomembr. 2007, 1768, 2764-2776). However, when a silica xerogel support was used, the phase behavior deviated from the phase diagram. Specifically, miscibility and trend lines of DSPC-rich domain area fraction, domain shape, and domain size versus cholesterol, obtained by analysis of fluorescence and atomic force microscopy (AFM) images, were as expected for mica-supported lipid bilayers, but were substantially stretched to higher cholesterol concentrations for silica xerogel-supported lipid bilayers. In addition, this behavior was found in three other ternary lipid compositions substituting slightly shorter acyl chain lengths in comparison to DSPC or a saturated lipid versus unsaturated DOPC. Qualitative comparison of domain characteristics of DOPC/DSPC/cholesterol (0 and 15 mol %) bilayers supported by silica xerogel, mica, borosilicate glass, and quartz showed that the networked surface layer of high curvature (0.04 nm(-1)) silica beads was the dominant influence as opposed to the surface chemistry. Based upon the literature, we postulate two curvature-based mechanisms that explain our results. In the first mechanism, cholesterol was transferred from the higher curvature supported lipid bilayer to the lower curvature vesicles in the medium during the vesicle fusion and thermal cooling step, resulting in a lowered cholesterol concentration of the supported lipid bilayer. In the second mechanism, high curvature promoted sustained lipid demixing as the cholesterol concentration was increased, thus creating a new phase diagram in which coexisting phases persist to a higher cholesterol concentration. These results suggest that a surface layer of high curvature features can be used to observe and study curvature-induced intrabilayer transport or demixing over large areas and that curvature can play an important role in sorting and localization of biomembrane components.
Department of Chemical Engineering & Materials Science, University of California, Davis, California 95616, USA.
Emel I
EI
Goksu
Marjorie L
ML
Longo
D002784:Cholesterol / Q000737:chemistry*; D008051:Lipid Bilayers; D018625:Microscopy, Atomic Force; D010713:Phosphatidylcholines / Q000737:chemistry; D012822:Silicon Dioxide
D008051:Lipid Bilayers; D010713:Phosphatidylcholines; D012822:Silicon Dioxide; D002784:Cholesterol; C017251:1,2-oleoylphosphatidylcholine