Purification and characterization of Rana pipiens brain Thy-1 glycoprotein.
The occurrence of Thy-1 antigens in Rana brain has been studied by the use of heterologous anti-Rana brain antisera raised in rabbit and BALB/c mouse (Thy-1.2) and AKR/J mouse (Thy-1.1) strains and by monoclonal anti-mouse Thy-1.1 and anti-mouse Thy-1.2 antibodies with the use of quantitative absorption assays. Three antigenic determinants were defined on Rana brain and referred to as: 1) the Rana-specific xenoantigen, 2) the Rana-mouse cross-reacting xenoantigen, and 3) the Thy-1.1 antigen. Thy-1 antigenic activities were solubilized from crude brain membranes in deoxycholate and followed by measuring the Rana-specific and the Thy-1.1 antigenic determinants. After solubilization, Rana brain Thy-1 antigens were purified by lentil lectin affinity chromatography and gel filtration on Sephadex G-200. A 605-fold and 400-fold enrichment in the Rana-specific and the Thy-1.1 antigenic activities with a yield of 25% and 17%, respectively, were obtained. Both antigenic activities were associated with a single glycoprotein of molecular size 3.1 nm and m.w. estimated at 27,000 by SDS-polyacrylamide gel electrophoresis. The serologic and biochemical properties of our purified Rana brain Thy-1 glycoprotein were very similar to those of the mammalian Thy-1 molecule, suggesting the conservation of the gene coding for Thy-1 during vertebrate evolution.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D000918:Antibody Specificity; D000954:Antigens, Surface / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D001666:Binding Sites, Antibody; D001921:Brain / Q000276:immunology*; D003429:Cross Reactions; D000939:Epitopes / Q000276:immunology / Q000302:isolation & purification; D005260:Female; D006023:Glycoproteins / Q000276:immunology / Q000302:isolation & purification*; D008099:Liver / Q000378:metabolism; D008297:Male; D051379:Mice; D008806:Mice, Inbred AKR; D008807:Mice, Inbred BALB C; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D011817:Rabbits; D011894:Rana pipiens / Q000276:immunology*; D018800:Thy-1 Antigens; D014982:Xenopus laevis
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D000939:Epitopes; D006023:Glycoproteins; D018800:Thy-1 Antigens