USE OF MONOCLONAL ANTI-LIGHT SUBUNIT ANTIBODIES TO STUDY THE STRUCTURE AND FUNCTION OF THE ENTAMOEBA-HISTOLYTICA GAL/GALNAC ADHERENCE LECTIN
USE OF MONOCLONAL ANTI-LIGHT SUBUNIT ANTIBODIES TO STUDY THE STRUCTURE AND FUNCTION OF THE ENTAMOEBA-HISTOLYTICA GAL/GALNAC ADHERENCE LECTIN
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DOI：
10.1007/bf00731279
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发表时间：
1994-10-01
影响因子：
3
通讯作者：
PETRI, WA
中科院分区：
文献类型：
--
作者：
MCCOY, JJ;WEAVER, AM;PETRI, WA
溶组织内阿米巴滋养体对宿主细胞的粘附是由半乳糖（Gal）和N-乙酰半乳糖胺（GalNAc）特异性表面凝集素介导的。凝集素是由通过二硫键连接的重（170 kDa）和轻（35-31 kDa）亚基组成的异二聚体蛋白。针对轻亚基-谷胱甘肽-S-转移酶融合蛋白的多克隆和单克隆抗体（mAb）被用来探测其结构和功能。产生了四种轻亚基特异性mAb，其识别活的不同轻亚基同种型上的不同表位。这些mAb的免疫印迹显示，当通过SDS-PAGE分析非还原滋养体蛋白时，轻亚基和重亚基共迁移，表明亚基不存在大量游离的异源二聚体。虽然抗重亚基抗体先前已被证明可以改变粘附性，但抗轻亚基抗体不能，这表明重亚基含有碳水化合物识别结构域。
Adherence of Entamoeba histolytica trophozoites to host cells is mediated by a galactose (Gal) and N-acetylgalactosamine (GalNAc)-specific surface lectin. The lectin is a heterodimeric protein composed of heavy (170 kDa) and light (35-31 kDa) subunits linked by disulfide bonds. Polyclonal and monoclonal antibodies (mAb) raised against a light subunit-glutathione-S-transferase fusion protein were used to probe its structure and function. Four light subunit-specific mAb were produced which recognized distinct epitopes on live different light subunit isoforms. Immunoblots with these mAb demonstrated co-migration of light and heavy subunits when nonreduced trophozoite proteins were analysed by SDS-PAGE, indicating that the subunits do not exist free of the heterodimer in significant quantities. While anti-heavy subunit antibodies had previously been shown to alter adherence, anti-light subunit antibodies did not, suggesting that the heavy subunit contains the carbohydrate recognition domain.