Preparation of tethered-type supported lipid bilayer using water-soluble silane coupling agent
JAPANESE JOURNAL OF APPLIED PHYSICS(2019)
摘要
Supported lipid bilayer (SLB) is one of the cell membrane model systems used to investigate the functions of lipids and membrane proteins. Tethered-type SLB (t-SLB) is a unique platform for controlling the distance and interaction between SLB and its substrate. In this study, we used a water-soluble silane coupling agent to modify a SiO2/Si substrate, keeping its surface roughness on the order of subnanometer, for the purpose of t-SLB fabrication. We modified the SiO2/Si substrate using carboxyethylsilanetriol (CEST) to terminate the surface with a carboxy group. The surface roughness was kept at similar to 0.15 nm (rms) after the CEST modification and following streptavidin conjugation. We prepared t-SLB that contained biotinylated lipids on the streptavidin-modified substrate using the vesicle fusion method and osmotic stimulation. Atomic force microscope observation and fluorescence recovery after photobleaching measurement showed that t-SLB was as flat as the SiO2/Si substrate and had approximately 20% smaller diffusion coefficient than non-tethered SLB. (C) 2019 The Japan Society of Applied Physics
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