Streptococcal erythrogenic toxin type C is not a phosphorylated protein. Description of two different purification procedures and investigation of its phosphorylation state

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 anti...

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Bibliographic Details
Published in:FEMS immunology and medical microbiology 1994-06, Vol.9 (1), p.65-76
Main Authors: Ozegowski, Jörg‐Hermann, Wollweber, Leo, Schmidt, Karl‐Hermann, Vettermann, Stefan, Reichardt, Werner, Köhler, Werner
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Animals
Antibodies, Monoclonal
Bacterial Proteins - immunology
Bacterial Proteins - isolation & purification
Bacteriology
Biological and medical sciences
Chromatography, Affinity
Chromatography, Agarose
Erythrogenic toxin type C
Exotoxins - immunology
Exotoxins - isolation & purification
Fundamental and applied biological sciences. Psychology
Humans
Lymphocyte Activation
Membrane Proteins
Mice
Mice, Inbred BALB C
Microbiology
Molecular Sequence Data
Pathogenicity, virulence, toxins, bacteriocins, pyrogens, host-bacteria relations, miscellaneous strains
Phosphorylation
Phosphorylation state
Streptococci
Streptococcus pyogenes - metabolism
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Summary: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.
ISSN:0928-8244
1574-695X
DOI:10.1111/j.1574-695X.1994.tb00475.x