The subunit structure of thymus leukemia antigens.
EDTA-containing buffer solubilizes thymus leukemia antigens (TLa) from crude thymocyte membrane fractions. The TL antigens consist mainly of molecules of a size similar to immunoglobulin G when gel chromatography analyses were performed under physiological conditions. A single component of TLa was apparent on sucrose density gradient ultracentrifugation of solubilized thymocyte membrane macromolecules as monitored by indirect immunoprecipitation. The sedimentation constant for the TL antigens (5.8 S) was considerably less than that for immunoglobulin G. The gel chromatography and ultracentrifugation data suggest an apparent molecular weight for TLa of about 120000. TLa isolated by indirect immunoprecipitation is composed of two types of polypeptide chains. The smaller subunit was identified as beta2-microglobulin. The larger polypeptide chain carried the alloantigenic determinants and displayed a molecular weight of about 50000 after reduction and alkylation. TLa subjected to molecular weight determination under denaturing conditions was composed of two components. The smaller component was beta2-microglobulin which evidently is linked to the larger polypeptide chain by noncovalent interactions only. The larger component had a size greater than reduced and alkylated immunoglobulin G heavy chains. Upon reduction and alkylation of the latter component its size was reduced and it appeared to have a molecular weight of about 50000. Consequently, TLa is composed of two disulfide linked heavy polypeptide chains and two beta2-microglobulin molecules. TLa solubilized by papain digestion comprises two polypeptide chains, one of which is beta2-microglobulin. The larger 37000-dalton subunit is a fragment of the heavy polypeptide chain. This was demonstrated by digesting solubilized 120000-dalton TLa with papain. The proteolytic fragments obtained were indistinguishable from those directly released from the cell surface by proteolysis. The papain-derived TLa fragment exhibited most if not all the alloantigenic determinants.
D000818:Animals; D000941:Antigens / Q000302:isolation & purification; D000951:Antigens, Neoplasm / Q000032:analysis*; D006649:Histocompatibility Antigens; D007942:Leukemia, Experimental / Q000276:immunology*; D051379:Mice; D008815:Mice, Inbred Strains; D008970:Molecular Weight; D010455:Peptides; D012995:Solubility; D013950:Thymus Gland / Q000032:analysis / Q000276:immunology*
D000941:Antigens; D000951:Antigens, Neoplasm; D006649:Histocompatibility Antigens; D010455:Peptides