Biochemical characterization including amino acid and carbohydrate compositions of canine and human brain Thy-1 antigen.
Human and canine brain Thy-1 antigens were solubilized in deoxycholate and antigen activity was followed both by conventional absorbed anti-brain xenosera of proven specificity and by mouse monoclonal antibodies to canine and human Thy-1. It is shown that greater than 80% of Thy-1 activity in the dog and man binds to lentil lectin, that the mobility on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis of canine and human Thy-1 is identical with that of rat Thy-1 and that the Stokes radius in deoxycholate of canine and human brain Thy-1 is 3.0 nm and 3.25 nm respectively. Both lentil lectin affinity chromatography followed by gel-filtration chromatography on the one hand and monoclonal antibody affinity chromatography on the other gave high degrees of purification of the brain Thy-1 molecule in the dog and man, resulting in single bands staining for both protein and carbohydrate on sodium dodecyl sulphate/polyacrylamide-gel electrophoresis (except for a slight contaminant of higher molecular weight staining for protein but not carbohydrate with human Thy-1 purified by lentil lectin and gel-filtration chromatography). Analysis of canine and human brain Thy-1 purified by monoclonal antibody affinity chromatography with additional gel filtration through Sephadex G-200 showed that these molecules had respectively 38% and 36% carbohydrate. The amino acid and carbohydrate compositions were similar to those previously reported for Thy-1 of the rat and mouse, the main point of interest being the presence in canine and human brain Thy-1 of N-acetylgalactosamine, which has been reported in rat and mouse brain Thy-1 but not in Thy-1 from other tissues.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000954:Antigens, Surface / Q000276:immunology; D001923:Brain Chemistry; D002241:Carbohydrates / Q000032:analysis; D002846:Chromatography, Affinity; D003840:Deoxycholic Acid; D004285:Dogs; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D037102:Lectins; D008565:Membrane Proteins / Q000276:immunology; D013045:Species Specificity; D018800:Thy-1 Antigens
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D002241:Carbohydrates; D037102:Lectins; D008565:Membrane Proteins; D018800:Thy-1 Antigens; D003840:Deoxycholic Acid