|Purified GST-tagged human Constitutive androstane receptor Hsp90 complex
GST-tagged Constitutive androstane receptor (CAR) consists of a 218 a.a. GST tag (Schistosoma japonicum Glutathione S-Transferase, Acc# 1M99_A, a 13 a.a. flexible linker and 352 a.a. human CAR, Acc# NP_001070948 (details). Constitutive androstane receptor is also known as nuclear receptor subfamily 1, group I, member 3 (NR1I3). In the cytoplasm it is found in complexes with Hsp90 which suggests its involvement in the receptor function (Yoshinari et al., FEBS Lett. 31;548(1-3):17-20, 2003) (details). It was produced in complex with human Hsp90 to facilitate native protein conformation and advanced performance in ligand binding and coactivator peptide recruitment assays.
Human Constitutive androstane receptor ligand-binding domain (CAR) in complex with human Hsp90 was produced in insect cells co-infected with two recombinant baculoviruses, one which encodes GST-tagged human Constitutive androstane receptor and another FoldHelper™-905c that provides for expression of Hsp40, Hsc70, Hsp90, Hop and p23 human molecular chaperones. Cells were cultivated in the absence of exogeneous ligands to ensure availability of the ligand-binding site. CAR/Hsp90 complex was purified from soluble cell fraction without the use of detergents, high-salt buffers or chaotropic agents and was formulated at about physiological conditions. GST tagged CAR and Hsp90 are the major components of the complex (Fig. 1).
20 mM Na-P pH 8.0, 40 mM KCl,
20 mM Na2MoO4, 7 mM reduced
glutathione, 15% glycerol.
Ligand binding and coactivator
peptide recruitment assays.
Handling upon arrival
Immediately store at -80°C.
Avoid freeze-thaw cycles.
While working, please keep
sample on ice.
Fig. 1. SDS-PAGE analysis of purified human Constitutive
androstane receptor in complex with Hsp90.
Proteins were separated in 4-20% SDS-PAGE and stained with
1. GST tagged Constitutive androstane receptor in
complex with Hsp90 and other molecular chaperones.
2. Low range molecular weight marker (Bio-Rad).