Collection and characterisation of bacterial membrane proteins

Massoud Saidijam, Georgios Psakis, Joanne L Clough, Johan Meuller, Shun'ichi Suzuki, Christopher J Hoyle, Sarah L Palmer, Scott M Morrison, Martin K Pos, Richard C Essenberg, Martin C J Maiden, Atif Abu-bakr, Simon G Baumberg, Alex A Neyfakh, Jeffrey K Griffith, Michael J Stark, Alison Ward, John O'Reilly, Nicholas G Rutherford, Mary K Phillips-JonesPeter J F Henderson, Gunnar von Heijne, Jan Rydstrom, Pater Brzezinski

    Research output: Contribution to journalArticlepeer-review

    30 Citations (Scopus)

    Abstract

    A general strategy for the amplified expression in Escherichia coli of membrane transport and receptor proteins from other bacteria is described. As an illustration we report the cloning of the putative a-ketoglutarate membrane transport gene from the genome of Helicobacter pylori, overexpression of the protein tagged with RGS(His)6 at the C-terminus, and its purification in mg quantities. The retention of structural and functional integrity was verified by circular dichroism spectroscopy and reconstitution of transport activity. This strategy for overexpression and purification is extended to additional membrane proteins from H. pylori and from other bacteria.

    Original languageEnglish
    Pages (from-to)170-5
    Number of pages6
    JournalFEBS Letters
    Volume555
    Issue number1
    DOIs
    Publication statusPublished - 27 Nov 2003

    Keywords

    • membrane transport protein
    • alpha-ketoglutarate
    • two-component system
    • Pathogen
    • heliobacter pylori

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