Reconstitution of Membrane Proteins into a Model Biological Membrane

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  • パターン化人工膜への膜タンパク質の再構成
  • パターンカ ジンコウマク エ ノ マク タンパクシツ ノ サイコウセイ

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Abstract

<p>Model biological membranes are a useful tool to study the molecular properties and functions of membrane proteins. We develop a strategy to directly reconstitute mammalian membrane proteins from the cell membrane into a model biological membrane to bypass the technically challenging solubilization and purification processes. We expressed dopamine D2 receptor (D2R), a G-protein coupled receptor (GPCR), in Chinese hamster ovary (CHO) cells and produced cell membrane blebs by chemical induction. By introducing blebs into a patterned framework of lithographically-polymerized lipid bilayer on the substrate surface, we could form a planar bilayer and observe single molecules of D2R. Interestingly, a much higher density of D2R molecules were reconstituted in a nanometric cleft between the substrate and a poly-dimethylsiloxane (PDMS) elastomer sheet. This methodology should enable to evaluate the physicochemical properties and functions of a wide range of mammalian membrane proteins.</p>

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