Cloning, expression, crystallization and preliminary X-ray diffraction studies of staphylococcal superantigen-like protein 1 (SSL1)

Acta Crystallogr F Struct Biol Commun. 2014 May;70(Pt 5):600-3. doi: 10.1107/S2053230X14006967. Epub 2014 Apr 15.

Abstract

Staphylococcus aureus produces a family of exotoxins which are structural homologues of superantigens and thus are called staphylococcal superantigen-like proteins (SSLs). Amongst the 14 SSL genes, ssl1 (SAOUHSC_00383) has been cloned in the pQE30 expression vector, overexpressed in Escherichia coli M15 (pREP4) cells and the protein purified to homogeneity. The protein was crystallized using 6% Tacsimate pH 6.0, 0.1 M MES pH 6.0, 25%(w/v) polyethylene glycol 3350, 100 mM NDSB 256 at 298 K by the sitting-drop vapour-diffusion method. The crystals belonged to space group P21, with unit-cell parameters a = 77.9, b = 70.5, c = 126.5 Å, β = 106.2°. X-ray diffraction data were collected and processed to a maximum resolution of 2.5 Å. The crystal contains six molecules in the asymmetric unit.

Keywords: SSL1; Staphylococcus aureus; superantigen-like proteins.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cloning, Molecular* / methods
  • Crystallization
  • Exotoxins / biosynthesis*
  • Exotoxins / chemistry
  • Exotoxins / genetics
  • Gene Expression Regulation, Bacterial*
  • Staphylococcus aureus* / genetics
  • Superantigens / biosynthesis*
  • Superantigens / chemistry
  • Superantigens / genetics
  • X-Ray Diffraction

Substances

  • Exotoxins
  • Superantigens