An analysis of the structure of the antigen receptor on a pigeon cytochrome c-specific T cell hybrid.
The monoclonal antibody A2B4-2 has been shown to bind to the antigen receptor on the cloned pigeon cytochrome c-specific T cell hybrid, 2B4. Initial immunoprecipitation and SDS-PAGE analysis with this clonotypic antibody demonstrated that the antigen receptor on this cell had a m.w. of 85,000 to 90,000. Under reducing conditions, the receptor protein appeared as two bands of 45,000 to 50,000 and 40,000 to 44,000 on an SDS-PAGE gel. In this paper the antigen receptor on this T cell hybrid is further characterized. The molecule is shown to be a heterodimer that exists in two different forms on the cell surface. Receptor molecules with an apparent m.w. of 84,000 and 86,000 were isolated by immunoprecipitation and separation on polyacrylamide gradient gels. After reduction, the individual alpha- and beta-chains were separated by isoelectric focusing. In both forms of the receptor, the acidic alpha-chain had an apparent m.w. of 42,000 to 44,000. This alpha-chain associated with one of two forms of beta-chain. One beta-chain had a m.w. of 42,000 to 44,000, with a pI range of 7.5 to 7.9, and the alternate form of the beta-chain, beta', had a m.w. of 46,000 to 48,000 and a more acidic pI range of 6.5 to 7.5. The results of this investigation indicate that under reducing conditions on SDS-PAGE gels, the original upper 45,000 to 50,000 m.w. band represented beta'-chains alone, whereas the lower 40,000 to 44,000 m.w. band represented a mixture of alpha- and beta-chains. Additional data are presented to indicate that this heterodimeric protein has intrachain as well as interchain disulfide bonds. This conclusion was reached from the characteristic pattern of protein migration on SDS-PAGE gels in the presence of a reducing agent concentration gradient. Thus, both chains of the antigen receptor must have intrachain disulfide bonds and may have similar domain structures.
Samelson
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D000818:Animals; D000911:Antibodies, Monoclonal; D001666:Binding Sites, Antibody; D055598:Chemical Phenomena; D002621:Chemistry; D010856:Columbidae / Q000276:immunology*; D003574:Cytochrome c Group / Q000276:immunology / Q000378:metabolism*; D004220:Disulfides / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000032:analysis; D006825:Hybridomas / Q000276:immunology / Q000378:metabolism*; D008970:Molecular Weight; D011948:Receptors, Antigen, T-Cell / Q000276:immunology / Q000302:isolation & purification* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D003574:Cytochrome c Group; D004220:Disulfides; D000939:Epitopes; D011948:Receptors, Antigen, T-Cell