Degradation of 2-phosphoglycerate by cytotoxin B of Clostridium difficile

Floyd Knoop, Robert Martig, Michaela Owens

Research output: Contribution to journalArticle

4 Scopus citations

Abstract

Cytotoxin B of C. difficile was highly purified by selective ammonium sulfate precipitation, Biogel A5m chromatography, phenyl boronate hydrophobic interaction chromatography and ultracentrifugation. The final cytotoxic product had a specific activity of 7.8 × 108 units mg protein and showed a single protein band with an estimated molecular weight of 163000 when subjected to SDS-PAGE. Immunoelectrophoresis of the final product showed a single precipitin arc. The addition of cytotoxin B to imidazole-HCl buffer (pH 7.4) containing MgSO4, KCl and the substrate 2-phosphoglycerate resulted in the formation of phosphoenolpyruvate as demonstrated by spectrophotometric analysis. Phosphoglycerate conversion was absent when the cytotoxin was heat-inactivated of reacted with specific antitoxin prior to assay.

Original languageEnglish (US)
Pages (from-to)9-12
Number of pages4
JournalFEBS Letters
Volume267
Issue number1
DOIs
StatePublished - Jul 2 1990

All Science Journal Classification (ASJC) codes

  • Biophysics
  • Structural Biology
  • Biochemistry
  • Molecular Biology
  • Genetics
  • Cell Biology

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