Molecular analysis of the Gal/GalNAc adhesin of Entamoeba histolytica
Molecular analysis of the Gal/GalNAc adhesin of Entamoeba histolytica
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DOI：
10.1111/j.1550-7408.1998.tb04518.x
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发表时间：
1998-03-01
影响因子：
2.2
通讯作者：
Lockhart, LA
中科院分区：
文献类型：
--
作者：
Mann, BJ;Lockhart, LA
半乳糖/N-乙酰-D-氨基半乳糖(Gal/GalNAc)可抑制的粘附素介导了溶组织内阿米巴与结肠上皮和其他多种靶细胞的附着。已有7种针对170 kDa亚基非重叠表位的单抗被证明可增强阿米巴粘附性。这7个单抗识别的表位被定位在170 kDa亚基胞外富含半胱氨酸的区域。通过检测在大肠杆菌中以融合蛋白形式表达的170 kDa亚基的分离区域以及变性的天然粘附素的单抗反应性来检验表位的构象性质。这些分析表明，3个单抗识别构象表位，其余4个识别线性表位。这些单抗的图谱已经确定了Gal/GalNAc粘附素的重要功能区域，并表明重组Gal/GalNAc粘附素在大肠杆菌中表达时，至少保留了一些天然构象。
Attachment of Entamoeba histolytica to colonic epithelium and a variety of other target cells is mediated by a galactose/N-acetyl D-galactosamine (Gal/GalNAc) inhibitable adhesin. Seven monoclonal antibodies specific for nonoverlapping epitopes of the 170 kDa subunit have been shown others enhance amebic adherence. The epitopes recognized by these seven monoclonal antibodies have been mapped to the extracellular cysteine rich region of the 170 kDa subunit. The conformational nature of the epitopes was examined by testing monoclonal antibody reactivity with isolated regions of the 170 kDa subunit expressed as fusion proteins in E. coli and also with denatured native adhesin. These analyses suggested that three of monoclonal antibodies recognized conformational epitopes while the remaining four recognized linear epitopes. The mapping of these monoclonal antibodies have identified functionally important regions of the Gal/GalNAc adhesin and have also shown that recombinant Gal/GalNAc adhesin, when expressed in E. coli, retained at least some of its native conformation.