Streptococcal erythrogenic toxin type C is not a phosphorylated protein. Description of two different purification procedures and investigation of its phosphorylation state
| Autor: | Jörg-Hermann Ozegowski, Werner Reichardt, Stefan Vettermann, Leo Wollweber, Karl-Hermann Schmidt, Werner Köhler |
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| Rok vydání: | 1994 |
| Předmět: |
Microbiology (medical)
Streptococcus pyogenes medicine.drug_class Molecular Sequence Data Immunology Erythrogenic toxin Exotoxins Biology Lymphocyte Activation Monoclonal antibody Microbiology Chromatography Affinity Sepharose Mice Phosphoamino Acids Bacterial Proteins Affinity chromatography medicine Animals Humans Immunology and Allergy Amino Acid Sequence Phosphorylation Mice Inbred BALB C medicine.diagnostic_test Elution Isoelectric focusing Antibodies Monoclonal Membrane Proteins General Medicine Chromatography Agarose Infectious Diseases Biochemistry Immunoassay |
| Zdroj: | FEMS Immunology and Medical Microbiology. 9:65-76 |
| ISSN: | 1574-695X 0928-8244 |
| DOI: | 10.1111/j.1574-695x.1994.tb00475.x |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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