Purification and characterization of mouse brain Thy-1.2 differentiation alloantigen.
Subcellular fractionation of C57BI/6J mouse brains produced a crude synaptosome preparation which contained virtually all of the Thy-1.2 antigenic activity of the isotonic whole brain homogenate. The Thy-1.2 was solubilized from the synaptosomes, following delipidation with acetone, by deoxycholate extraction. A glycoprotein fraction rich in Thy-1.2 was isolated from the bulk of the detergent-soluble material by lectin affinity chromatography. Fractionation of the lectin retentate by gel filtration chromatography produced a single peak of Thy-1.2 activity purified more than 2000-fold over the original homogenate. SDS polyacrylamide gel electrophoresis of this material revealed a single band which corresponded to an apparent molecular weight of 24,000. Amino acid composition data indicated that the protein portion of the molecule is similar to Thy-1.1 from mouse lymphoblastoid cells. Carbohydrate analysis revealed a qualitative similarity between mouse brain Thy-1.2 and Thy-1.1 from rat brain. Structural differences which could account for the Thy-1.1 and Thy-1.2 antigenic distinctions are apparently too subtle to be detected by compositional analysis.
D000596:Amino Acids / Q000302:isolation & purification; D000818:Animals; D001921:Brain / Q000276:immunology*; D002241:Carbohydrates / Q000302:isolation & purification; D002458:Cell Fractionation; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes; D007519:Isoantigens / Q000302:isolation & purification*; D051379:Mice; D008806:Mice, Inbred AKR; D008810:Mice, Inbred C57BL; D013569:Synapses / Q000276:immunology; D013574:Synaptosomes / Q000276:immunology
D000596:Amino Acids; D002241:Carbohydrates; D000939:Epitopes; D007519:Isoantigens