Cytosolic delivery and characterization of the TcdB glucosylating domain by using a heterologous protein fusion.

Abstract

TcdB from Clostridium difficile glucosylates small GTPases (Rho, Rac, and Cdc42) and is an important virulence factor in the human disease pseudomembranous colitis. In these experiments, in-frame genetic fusions between the genes for the 255 amino-terminal residues of anthrax toxin lethal factor (LFn) and the TcdB(1-556) coding region were constructed, expressed, and purified from Escherichia coli. LFnTcdB(1-556) was enzymatically active and glucosylated recombinant RhoA, Rac, Cdc42, and substrates from cell extracts. LFnTcdB(1-556) plus anthrax toxin protective antigen intoxicated cultured mammalian cells and caused actin reorganization and mouse lethality, all similar to those caused by wild-type TcdB.

• Authors: L M Spyres, M Qa'Dan, A Meader, J J Tomasek, E W Howard, J D Ballard
• Journal: Infection and immunity (Infect Immun) Vol. 69 Issue 1 Pg. 599-601 (Jan 2001) ISSN: 0019-9567 [Print] United States
• PMID: 11119561 (Publication Type: Journal Article)

Chemical References

• Antigens, Bacterial
• Bacterial Toxins
• Peptide Fragments
• Recombinant Fusion Proteins
• anthrax toxin
• GTP Phosphohydrolases

Topics

• Animals
• Antigens, Bacterial
• Bacterial Toxins (isolation & purification, metabolism)
• CHO Cells
• Clostridioides difficile (pathogenicity)
• Cricetinae
• Cytosol (metabolism)
• GTP Phosphohydrolases (metabolism)
• Glycosylation
• Humans
• Peptide Fragments (metabolism)
• Recombinant Fusion Proteins (metabolism)
• Substrate Specificity