A model system to study the insertion of cholesterol into a phospholipid monolayer.
Colloidal probe atomic force microscopy (AFM) was used to study the interaction between a surface bearing tethered cholesterol groups and an egg phosphatidylcholine (egg-PC) monolayer. The cholesterol bearing surface was comprised of a mixed self-assembled monolayer comprised of O-cholesteryl N-(8'-mecapto-3',6'-dioxaoctyl)carbamate (CPEO3) molecules and beta-mercaptoethanol formed on a 20 mum diameter gold-coated silica particle. The egg-PC monolayer was adsorbed onto an octadecylthiol monolayer formed on template-stripped gold. The force between the surfaces, as a function of separation, was measured for surface concentrations of CPEO3 from 0 to 100 mol %. At all concentrations there was a long-range repulsive double-layer force due to weak surface charges. At surface concentrations of CPEO3 from 1 to 29 mol % the interaction on the approach of the surfaces showed a maximum in the repulsive force, followed by a small (2-5 nm) jump into a force minimum corresponding to adhesion of the surfaces. On separation, a normalized pull-off force of 1.0-1.6 mN m(-1) was measured. Over the same concentration range, the calculated interaction energy per CPEO3 molecule decreased from 1.1 +/- 0.2 kT to 0.04 kT. At surface concentrations of 35 mol % and above there was no reproducible adhesion between the cholesterol-bearing surface and the phospholipid monolayer. We attribute the occurrence of short-range attraction and adhesion in the 1-29 mol % regime to the insertion of (some) cholesterol groups into the phospholipid monolayer. At higher surface concentrations the efficiency of insertion is reduced due to steric effects. We discuss the experimental results in the light of the energetics of the insertion of a cholesterol molecule into a lipid bilayer.
Sheikh
Khizar H
KH
School of Physics and Astronomy and Centre for Self-Organising Molecular Systems, The University of Leeds, Leeds LS2 9JT, United Kingdom.
Christenson
Hugo K
HK
Bushby
Richard J
RJ
Evans
Stephen D
SD
D002784:Cholesterol / Q000737:chemistry*; D008567:Membranes, Artificial; D018625:Microscopy, Atomic Force; D008956:Models, Chemical; D008958:Models, Molecular; D010743:Phospholipids / Q000737:chemistry*; D013499:Surface Properties
D008567:Membranes, Artificial; D010743:Phospholipids; D002784:Cholesterol