Pseudomonas aeruginosa exotoxin: purification by preparative polyacrylamide gel electrophoresis and the development of a highly specific antitoxin serum

Callahan, L T

doi:10.1128/iai.14.1.55-61.1976

Cited by 49 publications

(24 citation statements)

References 21 publications

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“…Antitoxin and anti-bovine serum albumin (BSA) were prepared in male New Zealand rabbits weighing 3 to 5 kg. The immunization schedule for preparation of antitoxin has been described previously (5). Anti-BSA serum was prepared by two intravenous injections of 4 mg of BSA (Miles Laboratories, Inc., Kankakee, I1l.)…”

Section: Methodsmentioning

confidence: 99%

“…Despite this accumulating information about the biological activities of exotoxin and the demonstration that pseudomonas infection in humans induces an antibody response to exotoxin A (19), its role in infection is unclear. In addition, although exotoxin is neutralized by antitoxin in vitro (5), the protective effect of antitoxin in experimental infection has not been adequately investigated. Although Liu and Hsieh (11) demonstrated protection in mice injected intraperitoneally with antitoxin against lethal doses of live P. aeruginosa, because of the relatively low virulence of this bacterium for normal mice, an unrealistically large inoculum was required to produce lethal disease in this model.…”

mentioning

confidence: 99%

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Passive protection by antitoxin in experimental Pseudomonas aeruginosa burn infections

Pavlovskis¹,

Pollack²,

Callahan³

et al. 1977

Infect Immun

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113

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The protective effect of intravenously administred rabbit antitoxin serum was studied in lethal Pseudomonas aeruginosa bum infections in mice. Survival after infection with 2 median lethal doses of a toxigenic, low-protease-producing strain (PA103) was enhanced in antitoxin-treated mice, as compared with controls that had received anti-bovine serum albumin serum (P = 0.0004). Survival time was prolonged in other antitoxin-treated mice infected with toxigenic, highprotease-producing strains (PA86 and PA220, P = 0.0003 and P = 0.01, respectively). In contrast, antitoxin had no protective effect in mice challenged with a nontoxigenic strain (WR 5, P = 0.57). There were fewer viable bacteria in blood and liver of antitoxin-treated mice than in those of anti-bovine serum albumintreated controls after infection with toxigenic organisms, whereas there were no significant differences between the two groups after challenge with the nontoxigenic strain. These data suggest that P. aeruginosa exotoxin A contributes to lethality in this burn infection model, and this effect is diminished by passive immunization with antitoxin.

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Passive protection by antitoxin in experimental Pseudomonas aeruginosa burn infections

Pavlovskis¹,

Pollack²,

Callahan³

et al. 1977

Infect Immun

Self Cite

113

View full text Add to dashboard Cite

show abstract

“…Previous studies have documented that purified Pseudomonas aeruginosa exotoxin A (2,3,9) inhibits protein synthesis in cultured mammalian cells (10,11) and in mouse organs (13). Iglewski and Kabat reported that toxin catalyzes the nicotinamide adenine dinucleotide (NAD)dependent inhibition of protein synthesis in rabbit reticulocyte lysates.…”

mentioning

confidence: 99%

Mechanism of action of Pseudomonas aeruginosa exotoxin A in experimental mouse infections: adenosine diphosphate ribosylation of elongation factor 2

Pavlovskis¹,

Iglewski²,

Pollack³

1978

Infect Immun

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The data presented indicate that one of the primary actions of Pseudomonas aeruginosa exotoxin during experimental infection is the inactivation of elongation factor 2 (EF-2) in various mouse organs. Organs from mice infected with the toxigenic P. aeruginosa strain PA103 contained considerably less EF-2 activity than did organs from uninfected controls. Whereas EF-2 activity was reduced in all organs examined from PA103-infected animals, the largest decrease was observed in the liver, where the active EF-2 levels were reduced by 70 to 90%. In addition, consistent inhibition of protein synthesis in livers but not in other organs was observed in mice infected with the toxigenic PA103 strain. Treatment of mice with antitoxin before infection with strain PA103 prevented inactivation of EF-2. When mice were infected with lethal doses of the nontoxigenic P. aeruginosa WR5 strain, tissue EF-2 levels were not markedly reduced below those derived from uninfected control animals.

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“…As the proteases released into culture supernatant might convert prohemolysin to the active form, it seemed important to reduce protease activity in the culture supernatant to identify the prohemolysin. Therefore, we added nitrilotriacetic acid (NTA) and ammonium sulfate to the culture medium; the former is a metal protease inhibitor (5), while the latter inhibits the production of proteases in various bacteria (22). The basal medium used was L broth.…”

Section: Production Of Precursor Of Hemolysinmentioning

confidence: 99%

Secretion of Hemolysin of Aeromonas sobria as Protoxin and Contribution of the Propeptide Region Removed from the Protoxin to the Proteolytic Stability of the Toxin

Nomura

Fujii

Okamoto

1999

Microbiology and Immunology

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The sequence at the amino terminus region of the hemolysin of Aeromonas sobria is homologous with that of aerolysin of A. hydrophila. However, there is no homology between the two toxins in the sequence at the carboxy terminal region. It has been shown that aerolysin is secreted into culture supernatant as aprotoxin. This proaerolysin is activated by the proteolytic removal of a carboxy terminal peptide. However, the role of the carboxy terminal region, which is removed in the activation process, has not been elucidated.In this study, we showed that hemolysin is also secreted as a protoxin into culture supernatant and that prohemolysin is cleaved by the protease of A. sobria between Ser-446 and Ala-447, resulting in the removal of a 42 amino acid peptide. The removal of the peptide converts the prohemolysin into active hemolysin. Subsequently, we mutated the hemolysin gene to delete the last several amino acid residues and expressed the genes in Escherichia coli, in order to examine the role of the carboxy terminal region of prohemolysin. The amounts of these mutant hemolysins accumulated in the periplasmic space of E. coli were very low compared with that of the wild-type. This observation indicated that the carboxy terminal region of prohemolysin contributes to the proteolytic stability of the toxin.

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Pseudomonas aeruginosa exotoxin: purification by preparative polyacrylamide gel electrophoresis and the development of a highly specific antitoxin serum

Cited by 49 publications

References 21 publications

Passive protection by antitoxin in experimental Pseudomonas aeruginosa burn infections

Passive protection by antitoxin in experimental Pseudomonas aeruginosa burn infections

Mechanism of action of Pseudomonas aeruginosa exotoxin A in experimental mouse infections: adenosine diphosphate ribosylation of elongation factor 2

Secretion of Hemolysin of Aeromonas sobria as Protoxin and Contribution of the Propeptide Region Removed from the Protoxin to the Proteolytic Stability of the Toxin

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