Cytosolic Delivery and Characterization of the TcdB Glucosylating Domain by Using a Heterologous Protein Fusion

AUTOR(ES)

Spyres, Lea M.

FONTE

American Society for Microbiology

RESUMO

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 TcdB1-556 coding region were constructed, expressed, and purified from Escherichia coli. LFnTcdB1-556 was enzymatically active and glucosylated recombinant RhoA, Rac, Cdc42, and substrates from cell extracts. LFnTcdB1-556 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.

Documentos Relacionados

• Multiplex PCR Targeting tpi (Triose Phosphate Isomerase), tcdA (Toxin A), and tcdB (Toxin B) Genes for Toxigenic Culture of Clostridium difficile
• I2B is a small cytosolic protein that participates in vacuole fusion
• Identification of a membrane fusion domain and an oligomerization domain in the baculovirus GP64 envelope fusion protein.
• Transcriptional activation of the glucose-regulated protein genes and their heterologous fusion genes by beta-mercaptoethanol.
• Genetic characterization of a mammalian protein-protein interaction domain by using a yeast reverse two-hybrid system.