Quantitation of lectin binding sites in human colon mucins by use of peanut and wheat germ agglutinins.
We have developed a novel method for quantitation of lectin binding sites in mucins derived from colon tissues. Binding of peanut agglutinin and wheat germ agglutinin was measured in extracts from normal and malignant human colon epithelium. Binding of wheat germ agglutinin was used as an estimate of the total mucin present in the tissue extract. Peanut agglutinin was found to bind to mucin from normal colon, but at levels that may be difficult to appreciate by fluorescence microscopy. The yield of mucin extracted from colon cancer was more variable than that from normal colon, and the binding ratio of peanut agglutinin to wheat germ agglutinin was greater in extracts from tumors than in normal tissues. Our findings confirm the histological observation that peanut agglutinin binds more avidly to mucins from colon cancer than to those from normal colon. The finding of peanut agglutinin binding sites in mucins front normal colon was not expected. The quantitative technique may have detected small numbers of binding sites not readily appreciable by fluorescence microscopy. Alternatively, the chromatographic method for measuring lectin binding may be sufficiently sensitive to detect nonspecific binding of the lectin to terminal galactose residues other than the Thomsen-Friedenreich antigen.
D010367:Arachis; D001665:Binding Sites; D002845:Chromatography; D003106:Colon / Q000378:metabolism*; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D006801:Humans; D037102:Lectins / Q000378:metabolism*; D009077:Mucins / Q000378:metabolism*; D019887:Peanut Agglutinin; D037121:Plant Lectins; D011975:Receptors, Mitogen / Q000032:analysis*; D013861:Thiocyanates; D014909:Wheat Germ Agglutinins / Q000378:metabolism*
D005452:Fluoresceins; D037102:Lectins; D009077:Mucins; D019887:Peanut Agglutinin; D037121:Plant Lectins; D011975:Receptors, Mitogen; D013861:Thiocyanates; D014909:Wheat Germ Agglutinins; D016650:Fluorescein-5-isothiocyanate