Purification and chemical characterization of a cholera toxin-cross-reactive cytolytic enterotoxin produced by a human isolate of Aeromonas hydrophila

J. M. Rose, C. W. Houston, D. H. Coppenhaver, J. D. Dixon, A. Kurosky

Research output: Contribution to journalArticle

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Abstract

A bacterial protein toxin possessing hemolytic, enterotoxic, and cytotoxic activities as well as cross-reactivity to cholera toxin was purified from culture filtrates of a human diarrheal isolate of Aeromonas hydrophila (SSU). This cytolytic enterotoxin was purified by ammonium sulfate precipitation, hydrophobic chromatography using phenyl-Sepharose, anion-exchange chromatography on DEAE-Bio-Gel A, and size-exclusion high-performance liquid chromatography. The factor was a single polypeptide with an apparent molecular weight of 52,000 as determined by polyacrylamide gel electrophoresis. Automated amino acid sequence analysis confirmed that the toxin was a single chain and established a 25-residue N-terminal segment which was identical to that of aerolysin purified from culture supernatants of A. hydrophila isolate Ab65 originally obtained from rainbow trout as reported by Howard et al. (S.P. Howard, W.J. Garland, M.J. Green, and J.T. Buckley, J. Bacteriol. 169:2869-2871, 1987). However, the amino acid compositional analysis of the toxin produced by our human isolate (SSU) differed significantly from that of the Ab65 isolate. Taken together, these results strongly indicated that several toxic phenomena associated with A. hydrophila (SSU) culture filtrates, including hemolysis, cytotoxicity, and enterotoxicity as well as cross-reactivity to cholera toxin, all can occur on a single polypeptide. In addition, these results underline the fact that although aerolysin-related toxins isolated from culture filtrates of A. hydrophila are biologically similar, significant chemical and immunological differences may exist between toxins produced by individual isolates.

Original languageEnglish (US)
Pages (from-to)1165-1169
Number of pages5
JournalInfection and Immunity
Volume57
Issue number4
StatePublished - 1989

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Aeromonas hydrophila
Cholera Toxin
Enterotoxins
Chromatography
Bacterial Toxins
Peptides
Bacterial Proteins
Poisons
Oncorhynchus mykiss
Protein Sequence Analysis
Ammonium Sulfate
Hemolysis
Anions
Polyacrylamide Gel Electrophoresis
Molecular Weight
Gels
High Pressure Liquid Chromatography
Amino Acids
aerolysin

ASJC Scopus subject areas

  • Immunology

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Purification and chemical characterization of a cholera toxin-cross-reactive cytolytic enterotoxin produced by a human isolate of Aeromonas hydrophila. / Rose, J. M.; Houston, C. W.; Coppenhaver, D. H.; Dixon, J. D.; Kurosky, A.

In: Infection and Immunity, Vol. 57, No. 4, 1989, p. 1165-1169.

Research output: Contribution to journalArticle

Rose, J. M. ; Houston, C. W. ; Coppenhaver, D. H. ; Dixon, J. D. ; Kurosky, A. / Purification and chemical characterization of a cholera toxin-cross-reactive cytolytic enterotoxin produced by a human isolate of Aeromonas hydrophila. In: Infection and Immunity. 1989 ; Vol. 57, No. 4. pp. 1165-1169.
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