Purification of protease from a mixture of exfoliative toxin and newborn-mouse epidermis.
Although the role of exfoliative toxin in staphylococcal scalded-skin syndrome has been suggested to be that of a serine protease, it has not been demonstrated to show proteolytic activity. Our purpose was to purify a proteolytic enzyme from a mixture of exfoliative toxin and newborn-mouse epidermis. We used gel filtration and ion-exchange and hydroxyapatite chromatography with a high-pressure liquid chromatography system. A casein-hydrolyzing enzyme was isolated from the mixture. The molecular mass of the enzyme was confirmed to be 20 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Subcutaneous injection of the purified enzyme into newborn mice reproduced the epidermal splitting that is seen in staphylococcal scalded-skin syndrome. These results suggest that exfoliative toxin does not work as a protease itself but that some reaction between exfoliative toxin and an epidermal component(s) first produces a protease, after which epidermal splitting occurs.
Ninomiya
J
J
Department of Dermatology, Showa University Fujigaoka Hospital, 1-30 Fujigaoka, Aoba-ku, Yokohama, Kanagawa, 227-0043 Japan. ninomiya@med.showa-u.ac.jp
Ito
Y
Y
Takiuchi
I
I
D000818:Animals; D000831:Animals, Newborn; D001427:Bacterial Toxins / Q000032:analysis* / Q000633:toxicity; D010450:Endopeptidases / Q000302:isolation & purification* / Q000633:toxicity; D051379:Mice; D012867:Skin / Q000187:drug effects* / Q000201:enzymology; D013211:Staphylococcus aureus / Q000472:pathogenicity*
D001427:Bacterial Toxins; D010450:Endopeptidases