| Purification and Quality Control | The His-tag recombinant protein is purified by affinity chromatography in combination with FPLC columns. The purified Retinoid X Receptor alpha is greater than 90% homogeneous based on SDS-PAGE analysis. | | Unit Definition (Activity) | 1 unit equals 1 nanogram of purified protein. 1 unit is sufficient for a gel mobility shift assay in a 20 µl reaction; 100 units are sufficient for protein-protein interaction assays. | | Applications | RXRα can be applied in reconstituted in vitro transcription assays and protein-protein interactions assays. | | Formulation and Storage | The protein is in 20mM Tris-HCl pH7.9,100mM NaCl, 0.2mM EDTA, 1mM DTT and 20% glycerol. Stored at -70°C before use. Avoid repeated freeze thaw cycles. | | Synonym | FLJ00280; FLJ00318; FLJ16020; FLJ16733; MGC102720 and NR2B1. | | Protein Sequence | MDTKHFLPLD FSTQVNSSLT SPTGRGSMAA PSLHPSLGPG IGSPGQLHSP ISTLSSPING
MGPPFSVISS PMGPHSMSVP TTPTLGFSTG SPQLSSPMNP VSSSEDIKPP LGLNGVLKVP
AHPSGNMASF TKHICAICGD RSSGKHYGVY SCEGCKGFFK RTVRKDLTYT CRDNKDCLID
KRQRNRCQYC RYQKCLAMGM KREAVQEERQ RGKDRNENEV ESTSSANEDM PVERILEAEL
AVEPKTETYV EANMGLNPSS PNDPVTNICQ AADKQLFTLV EWAKRIPHFS ELPLDDQVIL
LRAGWNELLI ASFSHRSIAV KDGILLATGL HVHRNSAHSA GVGAIFDRVL TELVSKMRDM
QMDKTELGCL RAIVLFNPDS KGLSNPAEVE ALREKVYASL EAYCKHKYPE QPGRFAKLLL
RLPALRSIGL KCLEHLFFFK LIGDTPIDTF LMEMLEAPHQ MT | | Background | Nuclear receptors form the largest known family of transcription factors and have a crucial role in nearly all aspects of vertebrate development and adult physiology by transducing the effects of hormones into transcriptional responses (1). The family is defined by two domains: (a) the central, highly conserved, DNA-binding domain (DBD) of approx. 66 amino acids, and (b) the C-terminal, structurally conserved, ligand-binding domain (LBD) of approx. 250 amino acids (2, 3). In addition to binding to DNA and activating transcription in response to 9-cis retinoic acid, RXR forms heterodimers with the receptors for thyroid hormone (TR), retinoic acid (RAR), vitamin D (VDR), prostanoids (PPAR), and numerous orphan receptors (4). RXR acts as both activator and repressor of transcription (5). In the absence of hormone, RXR (homo- or heterodimer) interacts with SMRT (silencing mediator for retinoid and thyroid hormone receptors) and N-CoR (nuclear receptor corepressor) and represses transcription through recruitment of histone deacetylases (6, 7). In the presence of hormone, RXR interacts with a number of activators including the SRC-1 family (8), CBP/p300 (9), pCAF (10) and the TRAP complex (11) to target chromatin acetylation activate of transcription (12). | | References | 1. Mangelsdorf, D. J., et al., (1995) Cell 83, 835–839
2. Glass, C. K. (1994) Endocrinol. Rev. 15, 391–407
3. Moras, D., et al., (1998) Curr. Opin. Cell Biol. 10, 384-391
4. Kliewer S.A., et al., (1992) Nature 355, 446–449
5. Glass C.K., et al., (1989) Cell 59, 697-708
6. Nagy L., et al., (1997) Cell 89, 373-380
7. Heinzel, T., et al., (1997) Nature 387, 43-48
8. Onate, S. A., et al., (1995) Science 270, 1354-1357
9. Kamei, Y., et al., (1996) Cell 85, 403-414
10. Blanco, J.C., et al., (1998) Genes Dev. 12, 1638-1651
11. Fondell, J.D., et al., (1996) Proc. Natl. Acad. Sci. USA 93, 8329-8333
12. Wolffe, A., et al., (1997) Genes Cells 2, 291-302 |
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