| Purification and Quality Control | The His-tag recombinant protein is purified by affinity chromatography in combination with FPLC columns. The purified Glucocorticoid Receptor is greater than 90% homogeneous based on SDS-PAGE analysis. | | Unit Definition (Activity) | 1 unit equals 1 nanogram of purified protein. 20 units are sufficient for a gel-mobility shift assay and 100 units are sufficient for a protein-protein interactions assay. | | Applications | GR has been applied in DNA and protein-protein interactions assays. | | Formulation and Storage | The protein is in 20mM Tris-HCl pH7.9, 300mM NaCl, 0.2mM EDTA, 1mM DTT and 20% glycerol. Stored at -70°C before use. Avoid repeated freeze thaw cycles. | | Synonym | GCCR; GCR; GR; GRL and NR3C1. | | Protein Sequence | MDSKESLTPG REENPSSVLA QERGDVMDFY KTLRGGATVK VSASSPSLAV ASQSDSKQRR
LLVDFPKGSV SNAQQPDLSK AVSLSMGLYM GETETKVMGN DLGFPQQGQI SLSSGETDLK
LLEESIANLN RSTSVPENPK SSASTAVSAA PTEKEFPKTH SDVSSEQQHL KGQTGTNGGN
VKLYTTDQST FDILQDLEFS SGSPGKETNE SPWRSDLLID ENCLLSPLAG EDDSFLLEGN
SNEDCKPLIL PDTKPKIKDN GDLVLSSPSN VTLPQVKTEK EDFIELCTPG VIKQEKLGTV
YCQASFPGAN IIGNKMSAIS VHGVSTSGGQ MYHYDMNTAS LSQQQDQKPI FNVIPPIPVG
SENWNRCQGS GDDNLTSLGT LNFPGRTVFS NGYSSPSMRP DVSSPPSSSS TATTGPPPKL
CLVCSDEASG CHYGVLTCGS CKVFFKRAVE GQHNYLCAGR NDCIIDKIRR KNCPACRYRK
CLQAGMNLEA RKTKKKIKGI QQATTGVSQE TSENPGNKTI VPATLPQLTP TLVSLLEVIE
PEVLYAGYDS SVPDSTWRIM TTLNMLGGRQ VIAAVKWAKA IPGFRNLHLD DQMTLLQYSW
MFLMAFALGW RSYRQSSANL LCFAPDLIIN EQRMTLPCMY DQCKHMLYVS SELHRLQVSY
EEYLCMKTLL LLSSVPKDGL KSQELFDEIR MTYIKELGKA IVKREGNSSQ NWQRFYQLTK
LLDSMHEVVE NLLNYCFQTF LDKTMSIEFP EMLAEIITNQ IPKYSNGNIK KLLFHQK | | Background | Glucocorticoids are a vital class of steroid hormones that mediate profound and diverse physiological effects in vertebrates from fish to man. Although named for their role in glucose homeostasis, glucocorticoids are eminently important throughout physiology, with regulatory roles in development, metabolism, neurobiology, programmed cell death, and many other functions. In addition to these far-reaching physiological roles, corticosteroids are among the most widely prescribed class of drugs in the world. The physiological response and sensitivity to glucocorticoids varies among species, individuals, tissues, cell types, and even during the cell cycle (1, 2). Additionally, several pathological conditions lead to, or are a result, of glucocorticoid resistance or hypersensitivity (3). The ligand-activated GR also interacts with a multitude of transcription factors such as c-jun (4), nuclear factor-B (NF-B) (5), the TFIID complex (6), STAT5 (7), as well as a host of coactivators (8). In addition, the GR interacts with numerous cytosolic proteins including chaperones, kinases, phosphatases, nuclear shuttling proteins, and the proteasome (9). | | References | 1. Hsu et al., (1995) J. Biol. Chem. 270, 3359-3364
2. Lim-Tio et al., (1997) Endocrinology 138, 2537-2543
3. Kino et al., (2001) Endocrinology 169, 437-445
4. Yangyen et al., (1990) Cell 62, 1205-1215
5. McKay et al., (1999) Endocr. Rev. 20, 435-459
6. Ford et al., (1997) Mol Endocrinol. 11, 1467-1475
7. Stocklin et al., (1996) Nature 383, 726-728
8. Jenkins et al., (2001) Endocrinol. Metab. 12, 122-126
9. Yudt et al., (2002) Mol. Endocrinol. 16, 1719-1726 |
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