title: Streptococcal erythrogenic toxin type C is not a phosphorylated protein. Description of two different purification procedures and investigation of its phosphorylation state.
abstract: Erythrogenic toxin type C (ETC) from different streptococcal group A strains was successively purified by absorption on phenylsepharose, acidic dialysis of the eluate at 40% saturated ammonium sulphate solution, CM-Sepharose chromatography, finally by immunoaffinity chromatography on monoclonal antibodies. Second, after growing of bacteria in the presence of [32P]orthophosphate to phosphorylate ETC, the ETC was purified with phenylsepharose following immunoaffinity chromatography. The occurrence of phosphoamino acids in the purified ETC was investigated by an immunoassay. No phosphoamino acids could be detected in the ETC molecule. Also after radiolabelling with 32P it was not possible to demonstrate a radioactive signal. The treatment with alkaline phosphatase has no influence on the mitogenicity or position of ETC in isoelectric focusing. The results obtained led to the conclusion that in contrast to the literature, ETC is not a phosphorylated protein.
authors.affiliation: Institut für Experimentelle Mikrobiologie der Friedrich-Schiller-Universität, Jena, FRG.
authors.forename: J H
authors.initials: JH
authors.lastname: Ozegowski
authors.forename: L
authors.initials: L
authors.lastname: Wollweber
authors.forename: K H
authors.initials: KH
authors.lastname: Schmidt
authors.forename: S
authors.initials: S
authors.lastname: Vettermann
authors.forename: W
authors.initials: W
authors.lastname: Reichardt
authors.forename: W
authors.initials: W
authors.lastname: Köhler
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins / Q000276:immunology / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D002847:Chromatography, Agarose; D005098:Exotoxins / Q000276:immunology / Q000302:isolation & purification*; D006801:Humans; D008213:Lymphocyte Activation; D008565:Membrane Proteins; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D010766:Phosphorylation; D013297:Streptococcus pyogenes / Q000378:metabolism*
chemical_list: D000911:Antibodies, Monoclonal; D001426:Bacterial Proteins; D005098:Exotoxins; D008565:Membrane Proteins; C117980:SpeA protein, Streptococcus pyogenes; C027951:erythrogenic toxin