Degradation of 2-phosphoglycerate by cytotoxin B of Clostridium difficile

Floyd C. Knoop; Robert Martig; Michaela Owens

doi:10.1016/0014-5793(90)80274-M

Degradation of 2-phosphoglycerate by cytotoxin B of Clostridium difficile

Floyd C. Knoop, Robert Martig, Michaela Owens

Department of Medical Microbiology and Immunology

Research output: Contribution to journal › Article

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 language	English (US)
Pages (from-to)	9-12
Number of pages	4
Journal	FEBS Letters
Volume	267
Issue number	1
DOIs	https://doi.org/10.1016/0014-5793(90)80274-M
Publication status	Published - Jul 2 1990

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All Science Journal Classification (ASJC) codes

Biochemistry
Biophysics
Molecular Biology

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Knoop, F. C., Martig, R., & Owens, M. (1990). Degradation of 2-phosphoglycerate by cytotoxin B of Clostridium difficile. FEBS Letters, 267(1), 9-12. https://doi.org/10.1016/0014-5793(90)80274-M

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10.1016/0014-5793(90)80274-M