Purification of Exfoliatin Produced by Staphylococcus aureus of Bacteriophage Group 2 and Its Physicochemical Properties

Abstract

An effective method for the isolation and purification of exfoliatin which has been recently reported by Melish and others as the staphylococcal toxin responsible for the scalded skin syndrome and the physicochemical properties of the purified toxin were described. From an active crude toxin produced by one of the clinical isolates of phage group 2, four types of toxic proteins which were all capable of causing the typical Nikolsky sign in neonatal mice were obtained and designated A, B, C, and D toxins. They had a molecular weight of about 24,000 and showed the same serological features in neutralization and precipitation tests, but were different from each other in showing a different single band with their respective mobilities in polyacrylamide disk electrophoresis. They were precipitated between pH 4.0 and 4.5 and lost their exfoliative capabilities. The resulting precipitates, however, could be solubilized in acetate buffer containing 0.5% sodium dodecyl sulfate, restoring their toxicities to almost the same extent as before. They were all stable when heated at 60 C for 60 min and at 100 C for 20 min, but lost their toxicities when heated at 100 C for 40 min. Additionally, the present authors observed that some staphylococcal strains not belonging to the typical phage group 2, isolated from patients with the scalded skin syndrome, were also capable of producing a similar but serologically unrelated exfoliative toxin. Recently several groups of workers have pointed out the close association between Staphylococcus aureus of phage group 2 and the disease of infants known as toxic epidermal necrolysis of the Ritter's type (3, 4, 7-9, 12, 13). An experimental model with newborn mice for the study of the toxic epidermal necrolysis (TEN) has been reported by Melish and Glasgow (10). Subsequently, Arbuthnott et al. (1)

Purification and Characterization of a novel Staphylococcus chromogenes Exfoliative Toxin

“…In 1994, we isolated a new exfoliative toxin (ETC) from a S. aureus strain isolated from a horse affected with phlegmon (Sato et al, 1994). ETA and ETB cause exfoliation in suckling mice (Kondo et al, 1973(Kondo et al, , 1974, but not in 1-day-old chicken whereas ETC causes exfoliation in both suckling mice and 1-day-old chickens (Sato et al, 1994). Recently, Yamaguchi et al (2002) isolated a new serotype of exfoliative toxin, ETD, from clinical isolates of S. aureus.…”

“…In contrast, SCET is a heat-stable toxin, as its toxicity is stable after being heated at 100°C for 20 min. It is well known that exfoliative toxin A (ETA) from S. aureus is a heat-stable toxin (Kondo et al, 1973). The toxicity of ETA is also stable after treatment at 100°C for 20 min and lost after being heated at 100°C for 40 min.…”

Partial Purification and Some Characteristics of Proteinase(s) Induced by Staphylococcal Exfoliative Toxin

“…Since Melish & Glasgow (1) demonstrated that staphylococcal scalded skin syndrome (SSSS) could be reproduced in newborn mice by injecting staphylococci cultured from a SSSS patient, it has been well established that intraepidermal splitting is caused by the action of staphylococcal exfoliative toxin (ET) (2)(3)(4). However, no precise mechanism of action has been elucidated.…”

An Exfoliative Toxin A‐Converting Phage Isolated from Staphylococcus aureus Strain ZM

“…S. aureus strain ZM, which was "a clinical isolate from a patient with SSSS (5), and derivatives of Terashima strain were used in this study (Table 1). Cultures for phage studies were prepared by overnight growth in oxoid nutrient broth (Unipath Ltd., Basingstoke, Hampshire, England) at 37 C. Phage particles arisen in the culture by the spontaneous lysis were separated from the bacterial cells using a high-speed centrifuge.…”