Retnoningrum D S, Podbielski A, Cleary P P
Department of Microbiology, University of Minnesota, Minneapolis 55455.
J Immunol. 1993 Mar 15;150(6):2332-40.
Streptococcus pyogenes, serotype M12, was discovered to express a human IgG3 binding protein. Western blot analysis of partially purified M12 protein, exposed to IgG3 myeloma protein, showed that both M12 Ag and the receptor protein were the same apparent size. A lambda clone (lambda 4.1) containing the emm12 open reading frame expressed both the M12 Ag and the IgG3 binding protein. The emm12 open reading frame was amplified by the polymerase chain reaction and subcloned into the expression vector pJLA602. Based on Western blot analysis, one recombinant Escherichia coli (pD3) expressed M12 protein with IgG3 binding activity. This result confirmed that the M12 protein from strain CS24 is also an IgG3 receptor. Deletion analyses showed that a truncated M12 protein encoded by an internal PvuII fragment was sufficient for IgG3 binding activity. Further deletion studies suggested that the IgG3 binding domain was located in a 200 amino acid internal fragment containing two directly repeated sequences. Other experiments suggest that the receptor did not bind to the same IgG3 domain as that recognized by protein G. The M12 protein did not bind human IgG1, IgG2, IgG4, or Ig from several other animal species.
化脓性链球菌M12血清型被发现可表达一种人IgG3结合蛋白。对部分纯化的M12蛋白与IgG3骨髓瘤蛋白进行的蛋白质印迹分析表明,M12抗原和受体蛋白的表观大小相同。一个包含emm12开放阅读框的λ克隆(λ4.1)表达了M12抗原和IgG3结合蛋白。通过聚合酶链反应扩增emm12开放阅读框,并亚克隆到表达载体pJLA602中。基于蛋白质印迹分析,一株重组大肠杆菌(pD3)表达了具有IgG3结合活性的M12蛋白。这一结果证实,来自CS24菌株的M12蛋白也是一种IgG3受体。缺失分析表明,由内部PvuII片段编码的截短M12蛋白足以具备IgG3结合活性。进一步的缺失研究表明,IgG3结合结构域位于一个包含两个直接重复序列的200个氨基酸的内部片段中。其他实验表明,该受体与蛋白G识别的IgG3结构域不同。M12蛋白不与人IgG1、IgG2、IgG4或其他几种动物物种的Ig结合。