Characterization of colon-specific antigen-p and isolation of immunologically active tryptic peptides.
CSAp was originally detected with antisera to an unfractionated extract of GW-39 human colonic carcinoma xenografts, and was determined to be restricted to normal and neoplastic gastrointestinal tissues and certain ovarian tumors. Gel filtration chromatography of GW-39 extract on Sepharose 4B columns reveals that more than 90% of the CSAp is associated with the void volume fraction. The smaller m.w. CSAp fraction is a population of fragments heterogeneous in respect to size but immunologically identical to the void CSAp. Efforts to dissociate intact CSAp from the void fraction by treatment with solutions of high ionic strength, SDS, non-ionic detergents, and 1 M lithium bromide have not been successful. CSAp is a glycoprotein that binds to Sepharose-concanavalin A and is distinct from other known tumor-associated antigens in immunologic and physicochemical properties. The antigen shows sensitivity to high concentrations of chaotropic reagents and especially to sulfhydryl reagents, even in low concentrations, which supports earlier results indicating that the CSAp antigenic determinant is associated with the polypeptide chain rather than with a carbohydrate moiety. CSAp has been reduced in size by either sonication or partial tryptic digestion. The former produces immunologically active fragments, heterogeneous in respect to size, that resemble the smaller size CSAp, whereas the tryptic digest generates 2 distinct peptides. The major tryptic peptide is found to have an approximate molecular size of 120,000, as determined by gel filtration.
D000818:Animals; D000941:Antigens; D001665:Binding Sites; D002850:Chromatography, Gel; D003106:Colon / Q000276:immunology*; D003208:Concanavalin A / Q000378:metabolism; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes; D010455:Peptides / Q000276:immunology* / Q000302:isolation & purification; D014357:Trypsin / Q000494:pharmacology
D000941:Antigens; D000939:Epitopes; D010455:Peptides; D003208:Concanavalin A; D014357:Trypsin