C型链球菌致热外毒素不是一种磷酸化蛋白。两种不同纯化方法的描述及其磷酸化状态的研究。

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

作者信息

Ozegowski J H, Wollweber L, Schmidt K H, Vettermann S, Reichardt W, Köhler W

机构信息

Institut für Experimentelle Mikrobiologie der Friedrich-Schiller-Universität, Jena, FRG.

出版信息

FEMS Immunol Med Microbiol. 1994 Jun;9(1):65-76. doi: 10.1111/j.1574-695X.1994.tb00475.x.

DOI:10.1111/j.1574-695X.1994.tb00475.x

PMID:7920465

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.

摘要

来自不同A组链球菌菌株的致热毒素C（ETC）先后通过在苯基琼脂糖上吸附、在40%饱和硫酸铵溶液中对洗脱液进行酸性透析、CM - 琼脂糖凝胶层析，最后通过在单克隆抗体上进行免疫亲和层析进行纯化。其次，在细菌于[32P]正磷酸盐存在下生长以使ETC磷酸化后，在免疫亲和层析之后用苯基琼脂糖对ETC进行纯化。通过免疫测定研究纯化的ETC中磷酸氨基酸的存在情况。在ETC分子中未检测到磷酸氨基酸。在用32P进行放射性标记后也无法显示放射性信号。用碱性磷酸酶处理对ETC在等电聚焦中的促有丝分裂活性或位置没有影响。所得结果得出结论，与文献相反，ETC不是一种磷酸化蛋白。