Characterization of a membrane-associated 3,3',5-triiodo-L-thyronine binding protein by use of monoclonal antibodies.
Four mouse hybridoma cell lines have been isolated which secrete antibodies to the membrane-associated thyroid hormone binding protein (Mr 55,000) from human epidermoid carcinoma A431 cells. J6 is rat specific; J2 is human and monkey specific; J8 and J9 have a wider specificity and react with similar thyroid hormone binding proteins (p55) from human, monkey, rat, and hamster. None of these antibodies reacts with mouse cells. J2, J6, and J9 are of the IgG1k class, and J8 is an IgAk antibody. p55 was characterized by using these monoclonal antibodies. It is not posttranslationally processed by glycosylation, phosphorylation, or sulfation. It has a cellular degradation rate t1/2 approximately equal to 3.2 h. Using immunofluorescence and electron microscopic immunocytochemistry, p55 was found to be associated with the lumenal face of the endoplasmic reticulum and nuclear envelope. When cell homogenates were prepared, significant amounts of p55 were released into the 110000g supernatant, indicating that p55 is loosely associated with the endoplasmic reticulum and nuclear envelope.
D000818:Animals; D000911:Antibodies, Monoclonal; D002352:Carrier Proteins / Q000302:isolation & purification*; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism* / Q000648:ultrastructure; D005455:Fluorescent Antibody Technique; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D008565:Membrane Proteins / Q000302:isolation & purification*; D051379:Mice; D008854:Microscopy, Electron; D009363:Neoplasm Proteins / Q000302:isolation & purification*; D011988:Receptors, Thyroid Hormone / Q000378:metabolism*; D013963:Thyroid Hormones; D014284:Triiodothyronine / Q000378:metabolism*; D000097810:Thyroid Hormone-Binding Proteins
D000911:Antibodies, Monoclonal; D002352:Carrier Proteins; D008565:Membrane Proteins; D009363:Neoplasm Proteins; D011988:Receptors, Thyroid Hormone; D013963:Thyroid Hormones; D014284:Triiodothyronine; D000097810:Thyroid Hormone-Binding Proteins