Characterization of rat gastric mucins using a monoclonal antibody, RGM23, recognizing surface mucous cell-type mucins.
A novel anti-mucin monoclonal antibody (mAb), designated RGM23, was developed against mucin purified from rat gastric mucosa. RGM23 reacted with the mucin attached to the ELISA well. The reactivity was lost by trypsin treatment, but not by periodate oxidation, indicating that RGM23 recognizes the peptide moiety of the mucin molecule. Histochemical study showed that RGM23 stained the corpus and antral surface mucosa of rat stomach, but not their glandular mucosa, nor duodenal, small intestinal or large intestinal mucosa. The area stained with RGM23 was coincident with that stained with 45M1, a mAb reacting with MUC5AC mucin. Examination of the mucin subunits extracted from rat stomach by Sepharose CL-4B and Q-Sepharose chromatography and CsTFA equilibrium centrifugation showed that RGM23 reacted with the surface mucous cell-type mucins that were stained with periodate-Schiff (PAS) and reacted with mAb RGM21. The gastric gland-type mucin, which reacted with mAb HIK1083, did not react with RGM23. On Q-Sepharose chromatography, a part of the RGM21-reactive mucins was only faintly stained with PAS and did not react with RGM23. The results together indicated that RGM23 probably reacted with the rat MUC5AC (rMuc5AC) mucin present in the surface mucosa of the stomach, and that the surface mucosal cells in rat stomach may contain mucin bearing non-rMuc5AC core protein in addition to rMuc5AC mucins.
Goso
Yukinobu
Y
Department of Biochemistry, Kitasato University Graduate School of Medical Sciences, 1-15-1 Kitasato, Sagamihara 228-8555. goso@kitasato-u.ac.jp
Ikezawa
Tomoaki
T
Kurihara
Makoto
M
Endo
Masaki
M
Hotta
Kyoko
K
Ishihara
Kazuhiko
K
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D002499:Centrifugation, Density Gradient; D002847:Chromatography, Agarose / Q000379:methods; D005752:Gastric Mucins / Q000737:chemistry* / Q000276:immunology; D005753:Gastric Mucosa / Q000737:chemistry* / Q000276:immunology; D007150:Immunohistochemistry; D055271:Mucin 5AC; D009077:Mucins / Q000032:analysis; D051381:Rats; D013270:Stomach / Q000737:chemistry* / Q000276:immunology; D014357:Trypsin
D000911:Antibodies, Monoclonal; D005752:Gastric Mucins; C524680:Muc5ac protein, rat; D055271:Mucin 5AC; D009077:Mucins; D014357:Trypsin