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J. Biol. Chem., Vol. 264, Issue 25, 14792-14796, 09, 1989
VM Gordon, WW Young Jr, SM Lechler, MC Gray, SH Leppla and EL Hewlett
Adenylate cyclase (AC) toxins produced by Bacillus anthracis and Bordetella
pertussis were compared for their ability to interact with and intoxicate
Chinese hamster ovary cells. At 30 degrees C, anthrax AC toxin exhibited a
lag of 10 min for measurable cAMP accumulation that was not seen with
pertussis AC toxin. This finding is consistent with previous data showing
inhibition of anthrax AC toxin but not pertussis AC toxin entry by
inhibitors of receptor-mediated endocytosis (Gordon, V. M., Leppla, S. H.,
and Hewlett, E. L. (1988) Infect. Immun. 56, 1066- 1069). Treatment of
target Chinese hamster ovary cells with trypsin or cycloheximide reduced
anthrax AC toxin-induced cAMP accumulation by greater than 90%, but was
without effect on pertussis AC toxin. In contrast, incubation of the AC
toxins with gangliosides prior to addition to target cells inhibited cAMP
accumulation by pertussis AC toxin, but not anthrax AC toxin. To evaluate
the role of lipids in the interaction of pertussis AC toxin with membranes,
multicompartmental liposomes were loaded with a fluorescent marker and
exposed to toxin. Pertussis AC toxin elicited marker release in a time- and
concentration- dependent manner and required a minimal calcium
concentration of 0.2 mM. These data demonstrate that the requirements for
intoxication by the AC toxins from B. anthracis and B. pertussis are
fundamentally different and provide a perspective for new approaches to
study the entry processes.
Adenylate cyclase toxins from Bacillus anthracis and Bordetella pertussis. Different processes for interaction with and entry into target cells
Department of Pharmacology and Medicine, University of Virginia, Charlottesville 22908.
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