The Qa-1 alloantigens. III. Biochemical analysis of the structure and extent of polymorphism of the Qa-1 allelic products.
Two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) was used to examine and compare the products of the Qa-1 locus. Analysis of Qa-1 isolated from detergent lysates of surface labeled cells indicated this molecule was a slightly acidic 48,000 to 50,000 dalton glycoprotein that displayed little charge heterogeneity on resting lymphocytes. The level of expression and degree of charge heterogeneity were both increased on activated lymphocytes. Direct comparison of the Qa-1b, Qa-1c, and Qa-1d allelic products by 2-D PAGE revealed that these three molecules could be distinguished from one another on the basis of isoelectric point, indicating that they were distinct at the molecular level. Comparison of Qa-1 isolated from several Qa-1b strains did not detect additional polymorphism. Removal of asparagine-linked oligosaccharides by treatment with endoglycosidase F indicated that carbohydrate contributed 10,000 to 12,000 to the m.w. of these allelic products. Comparative 2-D PAGE analysis could not distinguish between the deglycosylated Qa-1b, Qa-1c, and Qa-1d allelic products, implying that these molecules have similar primary structures. Peptide mapping supported this conclusion. Proteolytic digestion of the deglycosylated Qa-1b and Qa-1c allelic products resulted in identical peptide map patterns; such treatment of the deglycosylated Qa-1d allelic product produced a slightly different pattern. Peptide mapping analysis also demonstrated that the Tlaa and Qa-1a allelic products were distinct from one another, as well as being very different from the other three Qa-1 allelic products.
D000483:Alleles; D000818:Animals; D000918:Antibody Specificity; D015478:Antigens, Heterophile / Q000032:analysis; D000954:Antigens, Surface / Q000235:genetics* / Q000276:immunology; D002236:Carbohydrate Conformation; D004591:Electrophoresis, Polyacrylamide Gel; D005802:Genes, MHC Class II; D015395:Histocompatibility Antigens Class I; D007106:Immune Sera / Q000032:analysis; D007525:Isoelectric Focusing; D051379:Mice; D008805:Mice, Inbred A; D008810:Mice, Inbred C57BL; D008970:Molecular Weight; D010455:Peptides / Q000032:analysis; D011110:Polymorphism, Genetic
D015478:Antigens, Heterophile; D000954:Antigens, Surface; D015395:Histocompatibility Antigens Class I; D007106:Immune Sera; D010455:Peptides; C032907:Q surface antigens