Co-Investigator(Kenkyū-buntansha) |
OINUMA Tsutomu Miyazaki Medical College, Faculty of Medicine, Research Asscociate, 医学部, 助手 (20168842)
KAWANO Jun-ichi Miyazaki Medical College, Faculty of Medicine, Research Asscociate, 医学部, 助手 (10136822)
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Research Abstract |
Gastric mucus is thought to protect the gastric mucosa from mechanical trauma, desiccation, pathogenic microorganisms, acid and proteases. We purified Griffonia simplicifolia agglutinin (GSA)-II-binding mucus glycoprotein(GMG)from rat gastric mucosa by solubilization in a guanidine-containing buffer, gel permeation chromatography, Ricinus communis agglutinin(RCA)-I-affinity chromatography and GSA-II-affinity chromatography. Rat GMG showed high molecular weight on a Sephacryl S-1000 column, and a single band in 0.5%agarose-2% polyacrylamide composite gels and blots. A protein of about 60kDa was contained in the GMG preparation. GMG was deglycosylated with trifluoromethanesulfonic acid treatment. An antibody was rraised against deglycosylated GMG(deGMG). The antibody recognized deGMG, GMG, periodic acid-treated deGMG, and O-glycanase-digested dGMG, but did not react to trypsin-digested deGMG.These results that the antibody recognizes the peptide backbone of GMG.In imminohistochemistry, the mucous gel layr of the stomach luminal surface was stained with the antibody. The antibody recognized not only gastric mucous neck cells and pyloric gland cells but also gastric surface mucous cells, mucous cells in the duodenal gland, and goblet cells in the small intestine and colon. these indicate that GMG is a component of rat gastric mucus, and that the antibody recignizes mucous-secreting cells in rat stomach and intestine.
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