[WS4] APPA/PSSJワークショップ 蛋白質研究の世界連携に向けて

[WS4-05] Delivery of Recombinant SARS-CoV-2 Envelope Protein into Human Cells

James M. Hutchison^1,2, Ricardo Capone^2,3, Dustin D. Luu^4,5, Arina Hadziselimovic^2,3, Wade D. Van Horn^4,5, ^○Charles R. Sanders^2,3 (1.Chemical and Physical Biology Graduate Program, Vanderbilt University, 2.Center for Structural Biology, Vanderbilt University, 3.Department of Biochemistry, Vanderbilt University, 4.School of Molecular Sciences, Arizona State University, 5.The Biodesign Institute Centers for Personalized Diagnostics and Mechanisms of Evolution, Arizona State University)

The 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.