Peripheral alpha-linked N-acetylglucosamine on the carbohydrate moiety of mucin derived from mammalian gastric gland mucous cells: Epitope recognized by a newly characterized monoclonal antibody
Peripheral alpha-linked N-acetylglucosamine on the carbohydrate moiety of mucin derived from mammalian gastric gland mucous cells: Epitope recognized by a newly characterized monoclonal antibody
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DOI：
10.1042/bj3180409
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发表时间：
1996-09-01
影响因子：
4.1
通讯作者：
Hotta, K
中科院分区：
文献类型：
--
作者：
Ishihara, K;Kurihara, M;Hotta, K
为了获得一个工具来研究的结构特征和检测来自胃腺粘液细胞的粘蛋白，我们开发了一种单克隆抗体，命名为HIK 1083，从大鼠胃粘膜中纯化的粘蛋白。在ELISA中，HIK 1083与从胃体和胃窦的深层纯化的粘蛋白强烈反应，但仅与从表面粘膜层获得的粘蛋白轻微反应。用碱性硼氢化物还原抗原性粘蛋白得到的寡糖抑制了粘蛋白与HIK 1083的反应。根据核磁共振谱的证据，与抗原性粘蛋白单克隆抗体反应的两种纯化的寡糖糖醇分别具有一个和两个外周α-连接的N-乙酰葡萄糖胺残基。此外，β-单糖的市售对硝基苯基衍生物，只有对硝基苯基-N-乙酰基-α-D-氨基葡萄糖苷以浓度依赖性方式抑制该单克隆抗体与抗原性粘蛋白的反应。这些结果以及免疫组织化学观察结果表明，α-连接的N-乙酰葡糖胺残基特异性地附着于在胃腺型细胞中合成和分泌的粘蛋白的碳水化合物部分的外周区域，并表明单克隆抗体HIK 1083识别该结构。
To obtain a tool to study the structural characterization and the detection of mucin derived from the gastric gland mucous cells, we developed a monoclonal antibody, designated HIK1083, against mucin purified from rat gastric mucosa. In an ELISA, HIK1083 reacted strongly with the mucin purified from a deep layer of the corpus and antrum but only slightly reacted with that obtained from the surface mucosal layer. The reaction of mucin and HIK1083 was inhibited by the oligosaccharides obtained by the alkaline borohydride reduction of antigenic mucin, Two purified oligosaccharide alditols reacting with the monoclonal antibody obtained from the antigenic mucin had one and two peripheral alpha-linked N-acetylglucosamine residues, respectively, according to the evidence from NMR spectroscopy. Moreover, among-the commercially available p-nitrophenyl derivatives of monosaccharides, only p-nitrophenyl-N-acetyl-alpha-D-glucosaminide inhibited the reaction of this monoclonal antibody and the antigenic mucin in a concentration-dependent manner. These results, as well as the immunohistochemical observations, indicate that alpha-linked N-acetylglucosamine residues are specifically attached to the peripheral region of the carbohydrate moiety of the mucin synthesized in and secreted from the gastric-gland-type cells, and indicate that the monoclonal antibody HIK1083 recognizes this structure.