Crystallization and preliminary X-ray analysis of monalysin, a novel β-pore-forming toxin from the entomopathogen Pseudomonas entomophila

Acta Crystallogr Sect F Struct Biol Cryst Commun. 2013 Aug;69(Pt 8):930-3. doi: 10.1107/S174430911301885X. Epub 2013 Jul 27.

Abstract

Monalysin was recently described as a novel pore-forming toxin (PFT) secreted by the Drosophila pathogen Pseudomonas entomophila. Recombinant monalysin is multimeric in solution, whereas PFTs are supposed to be monomeric until target membrane association. Monalysin crystals were obtained by the hanging-drop vapour-diffusion method using PEG 8000 as precipitant. Preliminary X-ray diffraction analysis revealed that monalysin crystals belonged to the monoclinic space group C2, with unit-cell parameters a = 162.4, b = 146.2, c = 144.4 Å, β = 122.8°, and diffracted to 2.85 Å resolution using synchrotron radiation. Patterson self-rotation analysis and Matthews coefficient calculation indicate that the asymmetric unit contains nine copies of monalysin. Heavy-atom derivative data were collected and a Ta6Br14 cluster derivative data set confirmed the presence of ninefold noncrystallographic symmetry.

Keywords: Pseudomonas entomophila; monalysin; pore-forming toxins.

MeSH terms

  • Bacterial Toxins / chemistry*
  • Bacterial Toxins / isolation & purification
  • Crystallization
  • Crystallography, X-Ray
  • Pore Forming Cytotoxic Proteins / chemistry*
  • Pore Forming Cytotoxic Proteins / isolation & purification
  • Pseudomonas*

Substances

  • Bacterial Toxins
  • Pore Forming Cytotoxic Proteins