Adenylate cyclase toxins from Bacillus anthracis and Bordetella pertussis. Different processes for interaction with and entry into target cells.
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.
Gordon
V M
VM
Department of Pharmacology and Medicine, University of Virginia, Charlottesville 22908.
Young
W W
WW
Lechler
S M
SM
Gray
M C
MC
Leppla
S H
SH
Hewlett
E L
EL
D037361:Adenylate Cyclase Toxin; D000262:Adenylyl Cyclases / Q000378:metabolism* / Q000633:toxicity; D000818:Animals; D001408:Bacillus anthracis / Q000201:enzymology* / Q000378:metabolism; D001426:Bacterial Proteins / Q000378:metabolism / Q000633:toxicity; D002118:Calcium / Q000502:physiology; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D003513:Cycloheximide / Q000494:pharmacology; D005260:Female; D005732:Gangliosides / Q000378:metabolism; D007700:Kinetics; D008081:Liposomes; D010053:Ovary; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic / Q000378:metabolism; D010566:Virulence Factors, Bordetella / Q000378:metabolism* / Q000633:toxicity
D037361:Adenylate Cyclase Toxin; D001426:Bacterial Proteins; D005732:Gangliosides; D008081:Liposomes; D011956:Receptors, Cell Surface; D011971:Receptors, Immunologic; D010566:Virulence Factors, Bordetella; C034993:ganglioside receptor; D003513:Cycloheximide; D000262:Adenylyl Cyclases; D002118:Calcium