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Functional properties of human thyroid hormone receptor β1 overexpressed using baculovirus
Author(s) -
Collingwood T.N.,
Sydenham M.,
Page M.J.,
Chatterjee V.K.K.
Publication year - 1991
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(91)81310-5
Subject(s) - thyroid hormone receptor , recombinant dna , receptor , thyroid hormone receptor beta , microbiology and biotechnology , western blot , avidin , thyroid hormone receptor alpha , hormone , ligand (biochemistry) , thyroid , hormone receptor , biology , biotinylation , chemistry , nuclear receptor , biochemistry , endocrinology , transcription factor , gene , genetics , cancer , breast cancer
We have overexpressed the human β1 thyroid hormone receptor in insect cells using a recombinant baculovirus to a level of 5–10% of total cellular protein. The recombinant protein migrates as a 50 kDa band by SDS‐PAGE and Western blot analysis. The expressed receptor binds to L‐T3 with a K d of 1.3±0.4 × 10 −10 M and to thyroid hormone analogues with an affinity hierarchy of TRIAC>L‐T3>L‐T4>rT3. Gel retardation assays show highly specific receptor binding to a TRE which is modified by the presence of ligand and avidin—biotin complex DNA analysis shows a K d of 6.2±2.0 × 10 −10 M for this interaction. These results indicate high level expression of hTRβ with authentic hormone and DNA binding properties.