PT  - JOURNAL ARTICLE
AU  - James M. Hutchison
AU  - Ricardo Capone
AU  - Dustin D. Luu
AU  - Arina Hadziselimovic
AU  - Wade D. Van Horn
AU  - Charles R. Sanders
TI  - Delivery of recombinant SARS-CoV-2 envelope protein into human cells
AID  - 10.1101/2021.02.18.431684
DP  - 2021 Jan 01
TA  - bioRxiv
PG  - 2021.02.18.431684
4099  - http://biorxiv.org/content/early/2021/02/19/2021.02.18.431684.short
4100  - http://biorxiv.org/content/early/2021/02/19/2021.02.18.431684.full
AB  - 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 StatementThe authors have declared no competing interest.(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