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  • Targeting the expression of functional murine CMP-sialic acid transporter to the E. coli inner membrane

    Author(s)
    Maggioni, Andrea
    von Itzstein, Mark
    Gerardy-Schahn, Rita
    Tiralongo, Joe
    Griffith University Author(s)
    von Itzstein, Mark
    Maggioni, Andrea
    Tiralongo, Joe
    Year published
    2007
    Metadata
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    Abstract
    The heterologous expression of functional mammalian integral membrane proteins still represents a significant hurdle towards their crystallization and structure elucidation. We have therefore explored the use of the OmpA signal sequence to deliberately target the expression of the murine CMP-sialic acid transporter, a Golgi-resident protein with 10 putative transmembrane domains, to the Escherichia coli inner membrane. Here, we show that the expression of an OmpA signal sequence-FLAG-CMP-sialic acid transporter fusion protein in E. coli results in the targeting and insertion of recombinant protein within the inner membrane. ...
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    The heterologous expression of functional mammalian integral membrane proteins still represents a significant hurdle towards their crystallization and structure elucidation. We have therefore explored the use of the OmpA signal sequence to deliberately target the expression of the murine CMP-sialic acid transporter, a Golgi-resident protein with 10 putative transmembrane domains, to the Escherichia coli inner membrane. Here, we show that the expression of an OmpA signal sequence-FLAG-CMP-sialic acid transporter fusion protein in E. coli results in the targeting and insertion of recombinant protein within the inner membrane. Significantly, functionality was confirmed by the ability of spheroplasted E. coli and mixed phosphatidylcholine-E. coli inner membrane proteoliposomes incorporating recombinant CMP-sialic acid transporter to accumulate CMP-sialic acid in vitro.
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    Journal Title
    Biochemical and biophysical research communications
    Volume
    362
    Publisher URI
    http://www.elsevier.com/locate/ybbrc
    DOI
    https://doi.org/10.1016/j.bbrc.2007.08.070
    Copyright Statement
    © 2007 Elsevier. Please refer to the journal's website for access to the definitive, published version.
    Subject
    Medicinal and biomolecular chemistry
    Biochemistry and cell biology
    Medical biochemistry and metabolomics
    Publication URI
    http://hdl.handle.net/10072/18250
    Collection
    • Journal articles

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