CAR	Cat. #SC90

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).

Storage buffer 20 mM Na-P pH 8.0, 40 mM KCl, 20 mM Na2MoO4, 7 mM reduced glutathione, 15% glycerol. Purity >70% Applications 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   Coomassie blue.           1.        GST tagged Constitutive androstane receptor in                      complex with Hsp90 and other molecular chaperones.           2.        Low range molecular weight marker (Bio-Rad).