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J Biol Chem, Vol. 273, Issue 29, 18260-18267, July 17, 1998
From the Departments of Adenylate cyclase (AC) toxin from
Bordetella pertussis delivers its catalytic domain to the
interior of target cells where it converts host ATP to cAMP in a
process referred to as intoxication. This toxin also hemolyzes sheep
erythrocytes by a mechanism presumed to include pore formation and
osmotic lysis. Intoxication and hemolysis appear at strikingly
different toxin concentrations and evolve over different time scales,
suggesting that different molecular processes may be involved. The
present study was designed to test the hypothesis that intoxication and
hemolysis occur by distinct mechanisms.
Although the hemolytic activity of AC toxin has a lag of >1 h,
intoxication starts immediately. Because of this difference, we sought
a surrogate or precursor lesion that leads to hemolysis, and potassium
efflux has been observed from erythrocytes treated with other
pore-forming toxins. AC toxin elicits an increase in K+
efflux from sheep erythrocytes and Jurkat cells, a human T-cell leukemia line, that begins within minutes of toxin addition. The toxin
concentration dependence along with the analysis of the time course
suggest that toxin monomers are sufficient to elicit release of
K+ and to deliver the catalytic domain to the cell
interior. Hemolysis, on the other hand, is a highly cooperative event
that likely requires a subsequent oligomerization of these individual
units. Although induction of K+ efflux shares some
structural and environmental requirements with both intoxication and
hemolysis, it can occur under conditions in which intoxication is
reduced or prevented. The data presented here suggest that the
transmembrane pathway by which K+ is released is separate
and distinct from the structure required for intoxication but may be
related to, or a precursor of, that which is ultimately responsible for
hemolysis.
Distinct Mechanisms for K+ Efflux, Intoxication, and
Hemolysis by Bordetella pertussis AC Toxin
,
Medicine,
§ Molecular Physiology and Biological Physics,
¶ Pathology, and Pharmacology, University of Virginia
School of Medicine, Charlottesville, Virginia 22908
Copyright © 1998 by The American Society for Biochemistry and Molecular Biology, Inc.
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