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Recombinant SARS-CoV-2 envelope protein traffics to the trans-Golgi network following amphipol-mediated delivery into human cells.
Hutchison, James M; Capone, Ricardo; Luu, Dustin D; Shah, Karan H; Hadziselimovic, Arina; Van Horn, Wade D; Sanders, Charles R.
  • Hutchison JM; Chemical and Physical Biology Graduate Program, Vanderbilt University, Nashville, Tennessee, USA; Center for Structural Biology, Vanderbilt University, Nashville, Tennessee, USA.
  • Capone R; Center for Structural Biology, Vanderbilt University, Nashville, Tennessee, USA; Department of Biochemistry, Vanderbilt University, Nashville, Tennessee, USA.
  • Luu DD; School of Molecular Sciences, Arizona State University, Tempe, Arizona, USA; The Biodesign Institute Centers for Personalized Diagnostics and Mechanisms of Evolution, Arizona State University, Tempe, Arizona, USA.
  • Shah KH; School of Molecular Sciences, Arizona State University, Tempe, Arizona, USA; The Biodesign Institute Centers for Personalized Diagnostics and Mechanisms of Evolution, Arizona State University, Tempe, Arizona, USA.
  • Hadziselimovic A; Center for Structural Biology, Vanderbilt University, Nashville, Tennessee, USA; Department of Biochemistry, Vanderbilt University, Nashville, Tennessee, USA.
  • Van Horn WD; School of Molecular Sciences, Arizona State University, Tempe, Arizona, USA; The Biodesign Institute Centers for Personalized Diagnostics and Mechanisms of Evolution, Arizona State University, Tempe, Arizona, USA. Electronic address: wade.van.horn@asu.edu.
  • Sanders CR; Center for Structural Biology, Vanderbilt University, Nashville, Tennessee, USA; Department of Biochemistry, Vanderbilt University, Nashville, Tennessee, USA; Department of Medicine, Vanderbilt University Medical Center, Nashville, Tennessee, USA. Electronic address: chuck.sanders@vanderbilt.edu.
J Biol Chem ; 297(2): 100940, 2021 08.
Article in English | MEDLINE | ID: covidwho-1293905
ABSTRACT
The severe acute respiratory syndrome coronavirus 2 envelope protein (S2-E) is a conserved membrane protein that is important for coronavirus (CoV) assembly and budding. Here, we describe the recombinant expression and purification of S2-E in amphipol-class amphipathic polymer solutions, which solubilize and stabilize membrane proteins, but do not disrupt membranes. We found that amphipol delivery of S2-E to preformed planar bilayers results in spontaneous membrane integration and formation of viroporin cation channels. Amphipol delivery of the S2-E protein to human cells results in plasma membrane integration, followed by retrograde trafficking to the trans-Golgi network and accumulation in swollen perinuclear lysosomal-associated membrane protein 1-positive vesicles, likely lysosomes. CoV envelope proteins have previously been proposed to manipulate the luminal pH of the trans-Golgi network, which serves as an accumulation station for progeny CoV particles prior to cellular egress via lysosomes. Delivery of S2-E to cells will enable chemical biological approaches for future studies of severe acute respiratory syndrome coronavirus 2 pathogenesis and possibly even development of "Trojan horse" antiviral therapies. Finally, this work also establishes a paradigm for amphipol-mediated delivery of membrane proteins to cells.
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Full text: Available Collection: International databases Database: MEDLINE Main subject: Polymers / Propylamines / Surface-Active Agents / Cell Membrane / Trans-Golgi Network / Coronavirus Envelope Proteins Limits: Humans Language: English Journal: J Biol Chem Year: 2021 Document Type: Article Affiliation country: J.jbc.2021.100940

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Full text: Available Collection: International databases Database: MEDLINE Main subject: Polymers / Propylamines / Surface-Active Agents / Cell Membrane / Trans-Golgi Network / Coronavirus Envelope Proteins Limits: Humans Language: English Journal: J Biol Chem Year: 2021 Document Type: Article Affiliation country: J.jbc.2021.100940