Journal article
Characterization of Anthrolysin O, the Bacillus anthracis Cholesterol-Dependent Cytolysin
Infection and immunity, v 71(6), pp 3183-3189
Jun 2003
PMID: 12761097
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Abstract
We characterized the expression of a putative toxin of
Bacillus anthracis
, a member of the cholesterol-dependent cytolysin (CDC) family, which includes listeriolysin O, perfringolysin O, and streptolysin O. We named this cytotoxin anthrolysin O (ALO). Although
B. anthracis
expresses minimal hemolytic activity in clinical settings, we show that Sterne strain 7702 expresses hemolytic activity when grown in brain heart infusion broth or in other rich bacteriologic media, but it secretes barely detectable amounts of hemolysin when grown in Luria-Bertani (LB) broth. Glucose supplementation of LB broth increases the amount of secreted hemolytic activity. Expression of hemolytic activity is maximal during mid- to late-log phase and decreases in the stationary phase. These observations are supported, in part, by semiquantitative reverse transcriptase PCR of
alo
mRNA. Hemolytic activity in growth supernatants was increased in the presence of reducing agent and almost totally inhibited in a dose-dependent manner by cholesterol; both of these activities are characteristic of a CDC toxin. A mutant of Sterne strain 7702, strain UT231, in which the
alo
gene was deleted and replaced by a kanamycin cassette, secreted barely detectable hemolytic activity into the growth medium. When strain UT231 was complemented in
trans
with native
alo
on a low-copy-number plasmid [strain UT231(pUTE554)], it regained the ability to secrete hemolytic activity, indicating that ALO is the major hemolysin secreted by this strain of
B. anthracis
in rich media in vitro. To further support the
alo
gene product being a hemolysin, recombinant
B. anthracis
ALO (rALO) purified from
Escherichia coli
was extremely active against washed human erythrocytes, with complete hemolysis detected at ∼30 molecules of rALO per erythrocyte. Considering the virulence roles of CDCs for other gram-positive bacteria, we speculate that ALO may have a role in anthrax virulence.
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Details
- Title
- Characterization of Anthrolysin O, the Bacillus anthracis Cholesterol-Dependent Cytolysin
- Creators
- Jeffrey G Shannon - Department of Microbiology and Immunology, Drexel University College of Medicine, Philadelphia, Pennsylvania 19129Cana L Ross - Department of Microbiology and Immunology, Drexel University College of Medicine, Philadelphia, Pennsylvania 19129Theresa M Koehler - Department of Microbiology and Immunology, Drexel University College of Medicine, Philadelphia, Pennsylvania 19129Richard F Rest - Department of Microbiology and Immunology, Drexel University College of Medicine, Philadelphia, Pennsylvania 19129
- Publication Details
- Infection and immunity, v 71(6), pp 3183-3189
- Publisher
- American Society for Microbiology
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- [Retired Faculty]
- Web of Science ID
- WOS:000183116300025
- Scopus ID
- 2-s2.0-0037767296
- Other Identifier
- 991014878089104721
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- Collaboration types
- Domestic collaboration
- Web of Science research areas
- Immunology
- Infectious Diseases