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Purification and characterization of toxin B from Clostridium difficile
1Department of Microbiology and Immunology, University of Oklahoma Health Sciences Center, Oklahoma City 73190.
Infection and Immunity
|July 1, 1988
Summary
Clostridium difficile Toxin B was purified and characterized, revealing its molecular weight and amino-terminal sequence. This research provides essential data on the toxin
Area of Science:
- Microbiology
- Toxinology
Background:
- Clostridium difficile is a significant cause of antibiotic-associated diarrhea.
- Toxin B is a major virulence factor of C. difficile.
Purpose of the Study:
- To purify and characterize Toxin B from Clostridium difficile.
- To determine the molecular weight and amino-terminal sequence of Toxin B.
Main Methods:
- Purification using gel filtration and anion-exchange chromatography.
- Molecular weight determination via SDS-PAGE and gel filtration.
- Amino-terminal sequencing.
Main Results:
- Toxin B was purified to homogeneity.
- Estimated molecular weight of 250,000 (SDS-PAGE) and 500,000 (gel filtration), suggesting dimer formation.
- Homogeneity confirmed by SDS-PAGE, native PAGE, and chromatography.
- Amino-terminal sequence determined: NH3-Trp-Leu-Val-Asn-Arg-Lys-Gln-Leu-Glu-Lys-Met-Ala-Asn-Val-ARg-Phe-Arg.
Conclusions:
- The study successfully purified and characterized Clostridium difficile Toxin B.
- Molecular and sequence data provide a foundation for further toxicological studies.