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首页> 外文期刊>Infection and immunity >Characterization of group A streptococcal R-28 antigen purified by hydroxyapatite column chromatography.
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Characterization of group A streptococcal R-28 antigen purified by hydroxyapatite column chromatography.

机译：通过羟基磷灰石柱色谱纯化的A组链球菌R-28抗原的表征。

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Purified R-28 antigen from an M-protein-poor, R-antigen-rich strain of group A Streptococcus was prepared by sequential treatment of an acid extract of whole cells with ammonium sulfate fractionation and hydroxylapatite (HA) column chromatography. Purified R-28 antigen was eluted only with 0.10 M sodium phosphate, pH 6.7. Findings on quantitative amino acid composition, polyacrylamide gel electrophoresis pattern, and HA column elution pattern were similar but not identical to those previously reported for streptococcal M-proteins. Rabbits immunized with either HA-purified R-28 antigen or heat-killed cells developed two pepsin-sensitive, trypsin-resistant immunodiffusion lines of identity against HA-purified R-28 antigen but failed to form bactericidal antibody. One of these two lines formed a line of identity with R-28 antigen prepared by trypsinization of whole cells. The other line remained undefined, although it appeared not to be either streptococcal group A carbohydrate, M-protein, T-antigen, polyglycerophosphate, E4 antigen, or M-associated protein; by enzymatic criteria it is an R-antigen. Polyacrylamide gel electrophoresis of HA-purified R-28 antigen revealed multiple serologically active charge and size isomers. These findings suggest possible structural similarities between group A streptococcal M-proteins and R-antigens and also indicate that the same purification techniques may be utilized to study these protein antigens if the proper strain of Streptococcus is chosen.
机译：通过用硫酸铵分级分离和羟磷灰石（HA）柱色谱顺序处理全细胞的酸提取物，从A组链球菌的M蛋白贫乏，R抗原丰富的菌株中纯化R-28抗原。纯化的R-28抗原仅用0.10 M磷酸钠，pH 6.7洗脱。定量氨基酸组成，聚丙烯酰胺凝胶电泳图谱和HA柱洗脱图谱的发现与链球菌M蛋白的先前报道相似但不相同。用HA纯化的R-28抗原或热杀死的细胞免疫的兔子形成了两种针对HA纯化的R-28抗原的胃蛋白酶敏感，胰蛋白酶抗性免疫扩散谱系，但未能形成杀菌抗体。这两条系中的一条与通过全细胞胰蛋白酶消化制备的R-28抗原形成一条系。尽管似乎不是链球菌A组的碳水化合物，M蛋白，T抗原，聚甘油磷酸酯，E4抗原或M相关蛋白，但其他谱系仍不确定。按照酶学标准，它是一种R抗原。 HA纯化的R-28抗原的聚丙烯酰胺凝胶电泳显示出多种血清学活性电荷和大小异构体。这些发现表明A组链球菌M蛋白和R抗原之间可能的结构相似性，并且还表明如果选择合适的链球菌菌株，则可以使用相同的纯化技术来研究这些蛋白抗原。

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