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 α-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-α-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 α-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.
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September 1996
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Research Article|
September 01 1996
Peripheral α-linked N-acetylglucosamine on the carbohydrate moiety of mucin derived from mammalian gastric gland mucous cells: epitope recognized by a newly characterized monoclonal antibody
Kazuhiko ISHIHARA;
Kazuhiko ISHIHARA
‖
*Department of Chemistry, School of Medicine, Kitasato University, Sagamihara 228
‖To whom correspondence should be addressed.
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Makoto KURIHARA;
Makoto KURIHARA
†Department of Isehara Research Laboratory, Kanto Chemical Co. Inc., Isehara 259-11
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Yukinobu GOSO;
Yukinobu GOSO
‡Department of Biochemistry, School of Medicine, Kitasato University, Sagamihara 228
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Tsukiko URATA;
Tsukiko URATA
‡Department of Biochemistry, School of Medicine, Kitasato University, Sagamihara 228
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Hiroyoshi OTA;
Hiroyoshi OTA
§Central Research Laboratory, School of Medicine, Shinshu University, Matsumoto 390, Japan
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Tsutomu KATSUYAMA;
Tsutomu KATSUYAMA
§Central Research Laboratory, School of Medicine, Shinshu University, Matsumoto 390, Japan
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Kyoko HOTTA
Kyoko HOTTA
‡Department of Biochemistry, School of Medicine, Kitasato University, Sagamihara 228
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Publisher: Portland Press Ltd
Received:
October 06 1995
Revision Received:
April 11 1996
Accepted:
April 26 1996
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London © 1996
1996
Biochem J (1996) 318 (2): 409–416.
Article history
Received:
October 06 1995
Revision Received:
April 11 1996
Accepted:
April 26 1996
Citation
Kazuhiko ISHIHARA, Makoto KURIHARA, Yukinobu GOSO, Tsukiko URATA, Hiroyoshi OTA, Tsutomu KATSUYAMA, Kyoko HOTTA; Peripheral α-linked N-acetylglucosamine on the carbohydrate moiety of mucin derived from mammalian gastric gland mucous cells: epitope recognized by a newly characterized monoclonal antibody. Biochem J 1 September 1996; 318 (2): 409–416. doi: https://doi.org/10.1042/bj3180409
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