Biosynthesis and molecular nature of the T3 antigen of human T lymphocytes.
Immunoprecipitates of the T3 antigen prepared from HPB-ALL cells by using the monoclonal antibody UCH-T1 were analysed by SDS-polyacrylamide gel electrophoresis. Cells which had been biosynthetically labelled for up to 4 h gave a major polypeptide of mol. wt. 19 000 plus two weaker, more diffuse bands of mol. wts. 21 000 and 23 000, whereas surface labelled cells gave a prominent band of mol. wt. 19 000, a major band of 21 000 and a weaker diffuse band of approximately 26 000. As judged from their sensitivity to proteinase-K digestion, all the above polypeptides possess a transmembrane orientation. Digestion with endoglycosidases H and F (endo-H and endo-F), and tunicamycin treatment indicate that all the polypeptides, except that of 19 000 mol. wt. are N-glycosylated. The 21 000 and 23 000 mol. wt. chains possess both immature and mature oligosaccharide units, whereas the 26 000 mol. wt. band apparently has mature units only. Pulse chase experiments combined with digestion by endo-F and endo-H suggest that the N-glycosylated polypeptides are derived from two polypeptides of mol. wts. 14 000 and 16 000. It is concluded that the T3 antigen is derived from three different non-glycosylated polypeptides two of which are subsequently N-glycosylated to give the 21 000, 23 000 and 26 000 forms. The cell surface T3 antigen most probably comprises at least two distinct, non-covalently associated polypeptides, but the number and types of polypeptides giving rise to the whole molecule and whether different complexes exist is at present unclear.
D000911:Antibodies, Monoclonal; D000941:Antigens / Q000235:genetics* / Q000302:isolation & purification; D017252:CD3 Complex; D002460:Cell Line; D006020:Glycopeptides / Q000032:analysis; D006801:Humans; D007945:Leukemia, Lymphoid / Q000276:immunology*; D008970:Molecular Weight; D013601:T-Lymphocytes / Q000276:immunology*; D014415:Tunicamycin / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D000941:Antigens; D017252:CD3 Complex; D006020:Glycopeptides; D014415:Tunicamycin