Purification and characterization of a CHO cell-elongating toxin produced by Aeromonas hydrophila

A Chinese hamster ovary (CHO) cell-elongating toxin produced byAeromonas hydrophilawas purified from cell-free supernatant fluids by ammonium sulfate precipitation and fast protein liquid chromatography. The purified toxin had an isolelectric point (pI) of 3.7 and a molecular weight of 70000 in a si...

Full description

Saved in:
Bibliographic Details
Published in:Microbial pathogenesis 1995-07, Vol.19 (1), p.1-9
Main Authors: McCardell, Barbara A., Madden, Joseph M., Kothary, Mahendra H., Sathyamoorthy, Venugopal
Format: Article
Language:English
Subjects:
Aeromonas hydrophila
Aeromonas hydrophila - isolation & purification
Aeromonas hydrophila - metabolism
Amino Acid Sequence
Animals
Bacterial Toxins - chemistry
Bacterial Toxins - isolation & purification
Bacterial Toxins - pharmacology
Bacteriology
Biological and medical sciences
cell-elongating toxin
CHO Cells
Cricetinae
cytotonic toxin
Diarrhea - microbiology
effect on CHO cells
Female
Fundamental and applied biological sciences. Psychology
Humans
Intestinal Secretions - drug effects
Key words:Aeromonas hydrophila
Mice
Mice, Inbred ICR
Microbiology
Molecular Sequence Data
Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains
Pregnancy
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:A Chinese hamster ovary (CHO) cell-elongating toxin produced byAeromonas hydrophilawas purified from cell-free supernatant fluids by ammonium sulfate precipitation and fast protein liquid chromatography. The purified toxin had an isolelectric point (pI) of 3.7 and a molecular weight of 70000 in a single band on isoelectric focusing (IEF) gels and SDS-PAGE gels, respectively. The N-terminal sequence, amino acids 1–20, and the amino acid content were determined from Western blots of the 70 kDa band. No homology with any known microbial toxin was found. CHO cell activity was not neutralized by antiserum to cholera toxin (anti-CT), and the toxin did not react with anti-CT on Western blots. The toxin did not increase cyclic AMP, cyclic GMP, or prostaglandin E3levels in CHO cells. No cytotoxic activity was observed. Intragastric administration of purified toxin (5×104and 5×108CHO cell units) induced intestinal fluid accumulation in infant mice. These results suggest that this toxin may be a novel cytotonic toxin distinct from previously described toxins produced byA. hydrophilaorA. sobria.
ISSN:0882-4010
1096-1208
DOI:10.1006/mpat.1995.0039