The impact of clostridial glucosylating toxins on the proteome of colonic cells has been determined by analytical chemistry methods.
In a study published by Proteome Science, scientists used isotope-coded protein labelling techniques (ICPL) and mass spectrometry (LC-MALDI) to investigate whether the effect of certain toxins on target cells was dependant on catalytic glucosyltransferase activity.
The report explained that the anaerobe Clostridium difficile produces two major virulence factors - toxin A and B - that inactivate Rho proteins by glucosylation of a pivotal threonine residue.
These purified toxins trigger changes to the cytoskeleton and cell death in colonic cells.
Using analytical chemistry methods, the team compared the protein profiles of target cells that had been treated with wild type toxin A (rTcdA wt) or with a catalytically inactive mutant toxin A (mutant rTcdA).
As a result the scientists found that rTcdA wt impacted more cellular functions than just cytoskeleton reorganisation and apoptosis, with insight gained into the independent effects of clostridial glucosylating toxins on target cells after a short incubation time.
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