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Delivery of recombinant SARS-CoV-2 envelope protein into human cells
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Delivery of recombinant SARS-CoV-2 envelope protein into human cells
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Delivery of recombinant SARS-CoV-2 envelope protein into human cells
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Paper
Delivery of recombinant SARS-CoV-2 envelope protein into human cells
2021
Overview
Abstract SARS-CoV-2 envelope protein (S2-E) is a conserved membrane protein that is essential to coronavirus assembly and budding. Here, we describe the recombinant expression and purification of S2-E into amphipol-class amphipathic polymer solutions. The physical properties of amphipols underpin their ability to solubilize and stabilize membrane proteins without disrupting membranes. Amphipol delivery of S2-E to pre-formed planar bilayers results in spontaneous membrane integration and formation of viroporin ion channels. Amphipol delivery of the S2-E protein to human cells results in membrane integration followed by retrograde trafficking to a location adjacent to the endoplasmic reticulum-to-Golgi intermediate compartment (ERGIC) and the Golgi, which are the sites of coronavirus replication. Delivery of S2-E to cells enables both chemical biological approaches for future studies of SARS-CoV-2 pathogenesis and development of “Trojan Horse” anti-viral therapies. This work also establishes a paradigm for amphipol-mediated delivery of membrane proteins to cells. Competing Interest Statement The authors have declared no competing interest. * Abbreviations (SARS-CoV-2) Severe acute respiratory syndrome 2 virus (S2-E) SARS-CoV-2 envelope protein (ERGIC) endoplasmic reticulum-to-Golgi intermediate compartment (CoV) coronavirus (VLP) virus-like particle (ARDS) acute respiratory distress syndrome (NBD) nitrobenzoxadiazole (UPR) unfolded protein response
Publisher
Cold Spring Harbor Laboratory Press,Cold Spring Harbor Laboratory
