FUNCTIONAL-PROPERTIES OF HUMAN THYROID-HORMONE RECEPTOR-BETA-1 OVEREXPRESSED USING BACULOVIRUS

被引：7

作者：

COLLINGWOOD, TN

SYDENHAM, M

PAGE, MJ

CHATTERJEE, VKK

机构：

[1] UNIV CAMBRIDGE,ADDENBROOKES HOSP,DEPT MED,LEVEL 5,HILLS RD,CAMBRIDGE CB2 2QQ,ENGLAND

[2] WELLCOME FDN LTD,DEPT CELL BIOL,BECKENHAM,KENT,ENGLAND

来源：

FEBS LETTERS | 1991年 / 291卷 / 02期

基金：

英国惠康基金;

关键词：

THYROID HORMONE RECEPTOR; BACULOVIRUS; GENE EXPRESSION;

D O I：

10.1016/0014-5793(91)81310-5

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

We have overexpressed the human beta-1 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 x 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 x 10(-10) M for this interaction. These results indicate high level expression of hTR-beta with authentic hormone and DNA binding properties.

引用

页码：315 / 318

页数：4

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