Characterization of a chemically tritiated large molecular weight glucagon immunoreactive protein species by lectin-affinity column chromatography and reaction with anti-glucagon antibodies.
High molecular weight glucagon immunoreactive material, obtained by gel-filtration (in the presence of 6 M guanidine hydrochloride) of fetal bovine pancreatic extracts, was tritiated by reductive methylation. Concanavalin-A-Sepharose column chromatography of the radiolabeled preparation yielded a discrete Concanavalin-A-reactive, alpha-methyl-mannoside-displaceable radioactive peak, coinciding with the glucagon immunoreactive peak. Submission of the Con-A-reactive material to wheat germ agglutinin-Sepharose column chromatography yielded a lectin-reactive, N-acetyl-glucosamine-displaceable radioactive peak, coinciding with the glucagon immunoreactive peak. The tritiated Con-A-reactive component interacted specifically with anti-glucagon antibodies. Sephacryl S-200 gel-filtration (in the presence of guanidine hydrochloride) dissociated a approximately 40 kDa radioactive species from the antibody-antigen complex. These data provide direct evidence for the existence of a large molecular weight glycosylated glucagon-related protein species from the fetal bovine pancreas.
D000818:Animals; D000906:Antibodies; D000936:Antigen-Antibody Complex; D002417:Cattle; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D005333:Fetus; D005934:Glucagon / Q000302:isolation & purification*; D037102:Lectins; D008970:Molecular Weight; D010179:Pancreas / Q000032:analysis; D012685:Sepharose / Q000031:analogs & derivatives; D014316:Tritium; D014909:Wheat Germ Agglutinins
D000906:Antibodies; D000936:Antigen-Antibody Complex; D037102:Lectins; D014909:Wheat Germ Agglutinins; C028326:concanavalin A-sepharose; D014316:Tritium; D005934:Glucagon; D012685:Sepharose