Cytosolic delivery and characterization of the TcdB glucosylating domain by using a heterologous protein fusion.
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.
Spyres
L M
LM
The Department of Botany and Microbiology, University of Oklahoma, Norman, Oklahoma 73019-0245, USA.
Qa'Dan
M
M
Meader
A
A
Tomasek
J J
JJ
Howard
E W
EW
Ballard
J D
JD
D000818:Animals; D000942:Antigens, Bacterial; D001427:Bacterial Toxins / Q000302:isolation & purification / Q000378:metabolism*; D016466:CHO Cells; D016360:Clostridioides difficile / Q000472:pathogenicity*; D006224:Cricetinae; D003600:Cytosol / Q000378:metabolism*; D020558:GTP Phosphohydrolases / Q000378:metabolism; D006031:Glycosylation; D006801:Humans; D010446:Peptide Fragments / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D013379:Substrate Specificity
D000942:Antigens, Bacterial; D001427:Bacterial Toxins; D010446:Peptide Fragments; D011993:Recombinant Fusion Proteins; C030325:anthrax toxin; D020558:GTP Phosphohydrolases