Description

Viral protein U (Vpu) is a type-III integral membrane protein encoded by Human Immunodeficiency Virus-1 (HIV- 1). It is expressed in infected host cells and plays several roles in viral progeny escape from infected cells, including down-regulation of CD4 receptors.

Viral protein U (Vpu) is a type-III integral membrane protein encoded by Human Immunodeficiency Virus-1 (HIV- 1). It is expressed in infected host cells and plays several roles in viral progeny escape from infected cells, including down-regulation of CD4 receptors. But key structure/function questions remain regarding the mechanisms by which the Vpu protein contributes to HIV-1 pathogenesis. Here we describe expression of Vpu in bacteria, its purification and characterization. We report the successful expression of PelB-Vpu in Escherichia coli using the leader peptide pectate lyase B (PelB) from Erwinia carotovora. The protein was detergent extractable and could be isolated in a very pure form. We demonstrate that the PelB signal peptide successfully targets Vpu to the cell membranes and inserts it as a type I membrane protein. PelB-Vpu was biophysically characterized by circular dichroism and dynamic light scattering experiments and was shown to be an excellent candidate for elucidating structural models.

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    Title
    • Bacterial Expression, Correct Membrane Targeting, and Functional Folding of the HIV-1 Membrane Protein Vpu Using a Periplasmic Signal Peptide
    Date Created
    2017-02-22
    Resource Type
  • Text
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    Identifier
    • Digital object identifier: 10.1371/journal.pone.0172529
    • Identifier Type
      International standard serial number
      Identifier Value
      1045-3830
    • Identifier Type
      International standard serial number
      Identifier Value
      1939-1560
    Note
    • The article is published at http://journals.plos.org/plosone/article?id=10.1371/journal.pone.0172529

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    Deb, A., Johnson, W. A., Kline, A. P., Scott, B. J., Meador, L. R., Srinivas, D., . . . Mor, T. S. (2017). Bacterial expression, correct membrane targeting and functional folding of the HIV-1 membrane protein Vpu using a periplasmic signal peptide. Plos One, 12(2). doi:10.1371/journal.pone.0172529

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