Purification and chemical characterization of a cholera toxin-cross-reactive cytolytic enterotoxin produced by a human isolate of Aeromonas hydrophila.
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 Ah65 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 Ah65 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.
Rose
J M
JM
Department of Microbiology, University of Texas Medical Branch, Galveston 77550.
Houston
C W
CW
Coppenhaver
D H
DH
Dixon
J D
JD
Kurosky
A
A
D000333:Aeromonas / Q000276:immunology / Q000378:metabolism*; D000595:Amino Acid Sequence; D000818:Animals; D001427:Bacterial Toxins / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification*; D002460:Cell Line; D002772:Cholera Toxin / Q000276:immunology*; D006224:Cricetinae; D003412:Cricetulus; D003429:Cross Reactions; D003602:Cytotoxicity, Immunologic; D004768:Enterotoxins / Q000096:biosynthesis / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D008969:Molecular Sequence Data; D008970:Molecular Weight; D052899:Pore Forming Cytotoxic Proteins
D001427:Bacterial Toxins; D004768:Enterotoxins; D052899:Pore Forming Cytotoxic Proteins; C032676:aerolysin; D002772:Cholera Toxin