Construction and purification of His-tagged staphylococcal ArsB protein, an integral membrane protein that is involved in arsenical salt resistance.

Academic Article


  • Bacterial resistance to arsenical salts encoded on plasmid pI258 occurs by active extrusion of toxic oxyanions from cells of Staphylococcus aureus. The operon encodes for three gene products: ArsR, ArsB and ArsC. The gene product of arsB is an integral membrane protein and it is sufficient to provide resistance to arsenite and antimonite. A poly His-ArsB fusion protein was generated to purify the staphylococcal ArsB protein. Cells containing the His-tagged arsB gene were resistant to arsenite and antimonite. The levels of resistance to these toxic oxyanions by the His-tagged construct were greater than the levels obtained with the wild type gene. These data would indicate that the His-tagged protein is functionally active. A new 36 kDa protein band was visualized on 10% SDS-polyacrylamide gel electrophoresis (PAGE), which was confirmed as the His-ArsB protein by immunodetection with polyclonal Hisantibodies. The His-ArsB fusion protein was purified by the use of metal-chelate affinity chromatography with a Ni(+2)-nitrilotriacetic acid column and size-exclusion chromatography suggests that the protein was a homodimer.
  • Authors

  • Mascio, Carmela
  • White, Donald J
  • Tisa, Louis
  • Status

    Publication Date

  • September 2009
  • Has Subject Area

    Published In


  • Affinity chromatography
  • Arsenic resistance
  • Arsenite
  • Membrane protein
  • Transport
  • Digital Object Identifier (doi)

    Start Page

  • 212
  • End Page

  • 218
  • Volume

  • 49
  • Issue

  • 3