Artificial lipid membranes are often investigated as a replica of the cell membrane in the form of supported lipid bilayers (SLBs). In SLBs, the phase state of a lipid bilayer strongly depends on the presence of molecules such as cholesterol, ceramide, and physical parameters such as temperature. Cholesterol is a key molecule of biological membranes and it exerts condensing effect on lipid bilayers. In this paper, we demonstrate the influence of excess cholesterol content on a supported lipid bilayer of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC) (fluid-phase) using spectroscopic ellipsometry (SE) and coarse-grained (CG) molecular dynamics (MD) simulations. The results show the condensation effect due to cholesterol addition up to 30% and interleaflet decoupling at excess cholesterol beyond 30%. SE results show the separation of individual leaflets of the bilayer and influence of cholesterol on the biophysical properties such as thickness and optical index. CG simulations were performed at different ratios of DOPC:cholesterol mixtures to explore cholesterol-driven bilayer properties and stability. The simulations displayed the accumulation of cholesterol molecules at the interface of the lower and upper leaflets of the bilayer, thus leading to undulations in the bilayer. This work reports the successful application of SE technique to study lipid–cholesterol interactions for the first time.
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The authors thank the DST-Inspire Faculty scheme from the Department of Science and Technology (DST) (DST/INSPIRE/04/2015/000207) for the funding of this work. The authors also thank DIAT for infrastructural support in implementing this project. SK thanks DIAT for a Ph.D. fellowship. The authors also thank Dr. Suwarna Datar for her support.
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Kamble, S., Patil, S., Kulkarni, M. et al. Interleaflet Decoupling in a Lipid Bilayer at Excess Cholesterol Probed by Spectroscopic Ellipsometry and Simulations. J Membrane Biol 253, 647–659 (2020). https://doi.org/10.1007/s00232-020-00156-9
- Lipid membranes
- Spectroscopic ellipsometry
- MARTINI force field
- Coarse-grained simulations