Reversible Lifting of Surface Supported Lipid Bilayers with a Membrane-Spanning Nonionic Triblock Copolymer.

Abstract

Neutron reflectometry was used to monitor structural variations in surface-supported dimyristoylphosphatidycholine (DMPC) bilayers induced by the addition of Triton X-100, a surfactant commonly used to aid solubilization of membrane proteins, and the coaddition of a membrane spanning nonionic amphiphilic triblock copolymer, (PEO117-PPO47-PEO117, Pluronic F98). Surfactant addition causes slight compression of the bilayer thickness and the creation of a distinct EO layer that increases the hydrophilic layer proximal to the supporting substrate (i.e., a water and EO gap between the lipid bilayer and quartz) to 6.8 ± 0.4 Å. Addition of the triblock copolymer into the DMPC:Triton X-100 bilayer increases the complexity of (broadens) the lipid phase transition, further compresses the bilayer, and continues to expand the proximal hydrophilic layer thickness. The observed structural changes are temperature dependent with transmembrane polymer insertion achieved at 37 °C, leading to a compressed membrane thickness of 39.2 ± 0.2 Å and proximal gap of 45.0 ± 0.2 Å. Temperature-driven exclusion of the polymer at 15 °C causes partitioning of the polymer into the proximal space generating a large hydrogel cushion 162 ± 16 Å thick. An intermediate gap width (10-27 Å) is achieved at room temperature (22-25 °C). The temperature-driven changes in the proximal hydrophilic gap dimensions are shown to be reversible, but thermal history causes variation in magnitude. Temperature-driven changes in polymer association with a supported lipid bilayer offer a facile means to reversibly control both the membrane characteristics as well as the separation between membrane and solid substrate.