No abstract

A procedure for the purification of a protein marker for the staphylococci isolated from toxic-shock syndrome patients has been developed. The purification procedure involves the removal of the toxic protein from culture supernatant fluids of toxic-shock syndrome associated Staphylococcus aureus strains FRI-1169 and FRI-1183 by batch absorption with CG-50 resin, ion-exchange chromatography on CM-Sepharose CL-6B, and gel permeation chromatography on Sephacryl S-200. The purified toxin is a simple protein with a molecular weight of 24 000 +/- 500 as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The isoelectric point of the major band is 7.0 as determined by isoelectric focusing in polyacrylamide gels. The TS-toxin's reactivity with its specific antibody is not affected by tryptic digestion at pH 8.0 but is slowly reduced by treatment with pepsin at pH 4.5. The TS-toxin consists of 188 amino acid residues. Serine was shown to be the NH2-terminal amino acid residue by end-group analysis. Initial studies indicated the protein was emetic; thus tentatively it was called staphylococcal enterotoxin F. In this paper it is called TS-toxin because the emetic action in monkeys has not been confirmed.

An optimal sensitivity plate method for examining large numbers of staphylococcal strains for production of the known enterotoxins (A-E) is presented. Small volumes of relatively concentrated enterotoxin are produced by the semi-solid agar, cellophane-over-agar, or sac culture techniques. Detection of the enterotoxin in the supernatant fluid is accomplished with the optimal sensitivity plate method. In this method small plastic petri dishes (50 mm) were used for a modified Ouchterlony of high sensitivity.

A third staphylococcal enterotoxin C (C3) has been identified, purified, and characterized. Staphylococcal enterotoxin C3 was identified from a Staphylococcus aureus isolate received from England. The purified toxin was determined by gel permeation chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis to be a simple protein with a molecular weight of 26,900. The isoelectric point of the major band was determined by isoelectric focusing in polyacrylamide gels to be 8.15. The reaction of enterotoxin C3 with its specific antibody was not affected by tryptic digestion at pH 8.0 or peptic digestion at pH 4.5. The enterotoxin C3 consisted of 236 amino acid residues. Serine was shown to be the NH2-terminal amino acid residue by end group analysis. The protein was highly emetic in cynomolgus monkeys both per os and intravenously.