Recognition of a 56 kDa protein in partially purified rat hepatic nuclear thyroid hormone receptor by anti-human c-erb A beta antibody.
Human beta thyroid hormone receptor (c-erb A beta protein) produced by an Escherichia coli expression system was purified by sequential column chromatography followed by electroelution from an electrophoresis gel and an antibody was prepared. The antibody recognized a 56 kDa protein band in a partially purified rat hepatic nuclear thyroid hormone receptor fraction on Western blotting. Although multiple bands appeared on Western blotting of crude rat hepatic receptor preparations, a 56 kDa band was the most prominent and preadsorption of the antibody by purified c-erb A protein resulted in almost complete disappearance of the 56 kDa band, indicating that the 56 kDa band was formed by a specific antigen-antibody interaction. Furthermore, the 56 kDa protein appeared to co-elute with 3, 5, 3'-triiodo-L-thyronine binding activity in hydroxylapatite, Sephacryl S-200, and DNA-cellulose column chromatography of rat hepatic nuclear receptor, and sequential column purification resulted in selective enrichment of the 56 kDa band. These results suggest that the 56 kDa protein may be the major component of the rat hepatic thyroid hormone receptor.
Ichikawa
K
K
Department of Geriatrics, Endocrinology and Metabolism, Shinshu University School of Medicine, Matsumoto, Japan.
Hashizume
K
K
Kobayashi
M
M
Nishii
Y
Y
Sakurai
A
A
Takeda
T
T
Suzuki
S
S
Yamada
T
T
D000818:Animals; D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D066246:ErbB Receptors; D004926:Escherichia coli / Q000378:metabolism; D006801:Humans; D008099:Liver / Q000737:chemistry*; D008970:Molecular Weight; D009687:Nuclear Proteins / Q000032:analysis*; D011518:Proto-Oncogene Proteins / Q000276:immunology*; D051381:Rats; D011988:Receptors, Thyroid Hormone / Q000737:chemistry* / Q000302:isolation & purification
D000911:Antibodies, Monoclonal; D009687:Nuclear Proteins; D011518:Proto-Oncogene Proteins; D011988:Receptors, Thyroid Hormone; D066246:ErbB Receptors