Purification and characterization of Bordetella bronchiseptica dermonecrotic toxin.
Dermonecrotic toxin produced by Bordetella bronchiseptica was purified by chromatography on DEAE Toyopearl 650M and on Bio-Gel HTP, gel filtration on Sephadex G-200, and subsequent chromatography on Bio-Gel HTP and on SP Toyopearl 650M. The purified toxin was homogeneous by sodium dodecyl sulfate polyacrylamide gel electrophoresis and high performance liquid chromatography. There was a 90-fold increase in the dermonecrotic titer per mg protein in guinea pigs and the recovery of activity was 17.6% of that of the original cell extract. The purified toxin is a single-chain protein with a molecular weight of 145,000 and an isolelectric point of 6.3-6.7. Its minimal necrotizing dose is approximately 0.4 ng. It was completely inactivated by heating for 20 min at 56 degrees C. It contained no endotoxin, carbohydrates, nucleic acids, or hemagglutinins.
Research Center for Veterinary Science, Kitasato Institute, Chiba, Japan.
Y
Y
Horiguchi
T
T
Nakai
K
K
Kume
D000371:Agglutination; D001427:Bacterial Toxins / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism / Q000633:toxicity; D001884:Bordetella / Q000032:analysis*; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D008970:Molecular Weight; D011503:Transglutaminases; D010566:Virulence Factors, Bordetella
D001427:Bacterial Toxins; D010566:Virulence Factors, Bordetella; C060969:dermonecrotic toxin, Bordetella; D011503:Transglutaminases