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

IF 2.1 4区生物学 Q3 BIOTECHNOLOGY & APPLIED MICROBIOLOGY

Biotechnology Letters Pub Date : 2025-09-25 DOI:10.1007/s10529-025-03654-9

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

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Abstract

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.

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跨膜蛋白抗原与重组杆状病毒出芽粒子膜融合入二氧化硅微球支撑的磷脂双层。

对于人工细胞膜的重建，通常选择二氧化硅微珠作为支架来加固脆弱的脂质膜；这些双层覆盖的小珠通常被称为球形支撑脂质双层膜（SS-BLMs）。利用磷脂酰胆碱（PC）和磷脂酰丝氨酸（PS）的混合物制备单层小囊泡（suv），制备SS-BLMs。荧光显微镜观察发现，用十八烷基罗丹明B氯（R18）标记的加州自拟多核多角体病毒（AcMNPV）出芽病毒（BVs）可以融合，并在SS-BLMs上均匀分布。我们还发现，当BVs表达重组跨膜蛋白时，g蛋白偶联受体（GPCR）如β2肾上腺素能受体和促肾上腺皮质激素释放激素受体1，免疫荧光显微镜可以在SS-BLMs上特异性地观察到该蛋白。这表明重组蛋白通过病毒粒子-脂质双分子层膜融合进入球形支撑脂质双分子层。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Biotechnology Letters 工程技术-生物工程与应用微生物

CiteScore

5.90

自引率

3.70%

发文量

108

审稿时长

1.2 months

期刊介绍： Biotechnology Letters is the world’s leading rapid-publication primary journal dedicated to biotechnology as a whole – that is to topics relating to actual or potential applications of biological reactions affected by microbial, plant or animal cells and biocatalysts derived from them. All relevant aspects of molecular biology, genetics and cell biochemistry, of process and reactor design, of pre- and post-treatment steps, and of manufacturing or service operations are therefore included. Contributions from industrial and academic laboratories are equally welcome. We also welcome contributions covering biotechnological aspects of regenerative medicine and biomaterials and also cancer biotechnology. Criteria for the acceptance of papers relate to our aim of publishing useful and informative results that will be of value to other workers in related fields. The emphasis is very much on novelty and immediacy in order to justify rapid publication of authors’ results. It should be noted, however, that we do not normally publish papers (but this is not absolute) that deal with unidentified consortia of microorganisms (e.g. as in activated sludge) as these results may not be easily reproducible in other laboratories. Papers describing the isolation and identification of microorganisms are not regarded as appropriate but such information can be appended as supporting information to a paper. Papers dealing with simple process development are usually considered to lack sufficient novelty or interest to warrant publication.