Degradation of 2-phosphoglycerate by cytotoxin B of Clostridium difficile

Floyd C. Knoop, Robert Martig, Michaela Owens

Research output: Contribution to journalArticle

4 Citations (Scopus)

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

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Clostridium
Clostridium difficile
Cytotoxins
Chromatography
Degradation
Antitoxins
Precipitins
Phosphoenolpyruvate
Immunoelectrophoresis
Ultracentrifugation
Ammonium Sulfate
Hydrophobic and Hydrophilic Interactions
Polyacrylamide Gel Electrophoresis
Assays
Buffers
Proteins
Hot Temperature
Molecular Weight
Molecular weight
Substrates

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Biophysics
  • Molecular Biology

Cite this

Degradation of 2-phosphoglycerate by cytotoxin B of Clostridium difficile. / Knoop, Floyd C.; Martig, Robert; Owens, Michaela.

In: FEBS Letters, Vol. 267, No. 1, 02.07.1990, p. 9-12.

Research output: Contribution to journalArticle

Knoop, Floyd C. ; Martig, Robert ; Owens, Michaela. / Degradation of 2-phosphoglycerate by cytotoxin B of Clostridium difficile. In: FEBS Letters. 1990 ; Vol. 267, No. 1. pp. 9-12.
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