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<p>For the reconstitution of artificial cell membranes, silica microbeads are often selected as scaffolds to reinforce fragile lipid membranes; these bilayer-covered beads are often called spherical supported lipid bilayer membranes (SS-BLMs). SS-BLMs were made through the adsorption of small unilamellar vesicles (SUVs) preformed from a mixture of phosphatidylcholine (PC) and phosphatidylserine (PS). Fluorescence microscopic observation showed that budded viruses (BVs) of <i>Autographa californica</i> multiple nucleopolyhedrovirus (AcMNPV) labeled with octadecyl rhodamine B chloride (R18) could fuse and be optimally distributed homogeneously over the SS-BLMs when using a combination of PC and PS. We also demonstrated that when BVs expressing the recombinant transmembrane protein, G-protein coupled receptor (GPCR), like β2 adrenergic receptor and corticotropin-releasing hormone receptor 1, the protein could be specifically visualized on SS-BLMs by immunofluorescence microscopy. This indicates that recombinant proteins were incorporated into the spherical supported lipid bilayers through virion-lipid bilayer membrane fusion.</p>

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Incorporation of transmembrane protein antigens into phospholipid bilayers supported on silica microbeads using membrane fusion with budded virions of recombinant baculovirus

  • 期刊论文

  • 发表日期：

    2025年9月25日

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  • Kohei Nakanishi,
  • Seiwa Nishio,
  • Yushi Isozaki,
  • Masahiro Tomita,
  • Kanta Tsumoto

摘要

For the reconstitution of artificial cell membranes, silica microbeads are often selected as scaffolds to reinforce fragile lipid membranes; these bilayer-covered beads are often called spherical supported lipid bilayer membranes (SS-BLMs). SS-BLMs were made through the adsorption of small unilamellar vesicles (SUVs) preformed from a mixture of phosphatidylcholine (PC) and phosphatidylserine (PS). Fluorescence microscopic observation showed that budded viruses (BVs) of Autographa californica multiple nucleopolyhedrovirus (AcMNPV) labeled with octadecyl rhodamine B chloride (R18) could fuse and be optimally distributed homogeneously over the SS-BLMs when using a combination of PC and PS. We also demonstrated that when BVs expressing the recombinant transmembrane protein, G-protein coupled receptor (GPCR), like β2 adrenergic receptor and corticotropin-releasing hormone receptor 1, the protein could be specifically visualized on SS-BLMs by immunofluorescence microscopy. This indicates that recombinant proteins were incorporated into the spherical supported lipid bilayers through virion-lipid bilayer membrane fusion.