Issue 2, 2022

Embedding a membrane protein into an enveloped artificial viral replica


Natural enveloped viruses, in which nucleocapsids are covered with lipid bilayers, contain membrane proteins on the outer surface that are involved in diverse functions, such as adhesion and infection of host cells. Previously, we constructed an enveloped artificial viral capsid through the complexation of cationic lipid bilayers onto an anionic artificial viral capsid self-assembled from β-annulus peptides. Here we demonstrate the embedding of the membrane protein Connexin-43 (Cx43), on the enveloped artificial viral capsid using a cell-free expression system. The expression of Cx43 in the presence of the enveloped artificial viral capsid was confirmed by western blot analysis. The embedding of Cx43 on the envelope was evaluated by detection via the anti-Cx43 antibody, using fluorescence correlation spectroscopy (FCS) and transmission electron microscopy (TEM). Interestingly, many spherical structures connected to each other were observed in TEM images of the Cx43-embedded enveloped viral replica. In addition, it was shown that fluorescent dyes could be selectively transported from Cx43-embedded enveloped viral replicas into Cx43-expressing HepG2 cells. This study provides a proof of concept for the creation of multimolecular crowding complexes, that is, an enveloped artificial viral replica embedded with membrane proteins.

Graphical abstract: Embedding a membrane protein into an enveloped artificial viral replica

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Article information

Article type
18 Aug 2021
20 Dec 2021
First published
21 Dec 2021
This article is Open Access
Creative Commons BY license

RSC Chem. Biol., 2022,3, 231-241

Embedding a membrane protein into an enveloped artificial viral replica

H. Furukawa, H. Inaba, Y. Sasaki, K. Akiyoshi and K. Matsuura, RSC Chem. Biol., 2022, 3, 231 DOI: 10.1039/D1CB00166C

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