共查询到20条相似文献,搜索用时 31 毫秒
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[3H]Triamcinolone acetonide was used to tag covalently specific glucocorticoid receptors by photoaffinity labelling at lambda greater than or equal to 320 nm. Receptors of wild-type mouse lymphoma cells and two glucocorticoid resistant mutants of "nuclear transfer deficient" (nt-) and "increased nuclear transfer" (nti) phenotypes, respectively, were used. Wild-type and nt- receptors yielded radiolabelled polypeptide bands of mol. wt. 98 000 as revealed by gel electrophoresis under denaturing conditions and fluorography. In contrast, the nti receptor had a mol. wt. of 42 000. Partial proteolysis of the wild-type receptor with alpha-chymotrypsin resulted in a fragment of mol. wt. 39 000 which still contained the steroid binding site but had increased affinity for DNA indistinguishable from that of the nti receptor. Chymotrypsin thus removed a domain from the wild-type receptor polypeptide which is involved in modulating DNA binding. The same domain is missing from the nti receptor. 相似文献
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The differences in the molecular properties of the nuclear aryl hydrocarbon (Ah) receptor from human Hep G2 and mouse Hepa 1c1c7 cells were investigated by time-dependent partial proteolysis with chymotrypsin or trypsin followed by column chromatographic and velocity sedimentation analysis. The sedimentation coefficients, Stokes radii and apparent molecular weights of the untreated human and mouse Ah receptor complexes were similar. Treatment of the nuclear Ah receptor complexes from both cell lines with chymotrypsin for 10 or 60 min gave lower molecular weight proteolytic products which also exhibited comparable molecular properties and salt gradient elution profiles from Sepharose columns linked to DNA. Treatment of the human and mouse nuclear Ah receptor complexes with trypsin (5 micrograms/mg protein) for 10 or 60 min gave a minor low molecular weight (29.7- or 25.7-kDa) proteolysis product which was detected only with the mouse Hepa 1c1c7 Ah receptor complex. The time- and concentration-dependent proteolytic digest maps of the human and mouse Ah receptor were determined using receptor preparations which were photoaffinity labeled with [125I]7-iodo-2, 3-dibromodibenzo-p-dioxin. The human Ah receptor was significantly more resistant to proteolysis by trypsin or chymotrypsin than the mouse Ah receptor. At a low concentration of chymotrypsin (1 microgram/mg protein) the Hepa 1c1c7 receptor was degraded to two lower molecular weight fragments with apparent M(r) values at 71- and 48-kDa whereas the Hep G2 Ah receptor was relatively stable under these conditions. Although the human Ah receptor was more slowly hydrolyzed than the mouse receptor by trypsin, the major photolabeled breakdown products for the Ah receptor from both cell lines were observed at M(r) 48- and 45-kDa. The results of this study demonstrate that there were subtle but significant differences in the human and mouse Ah receptor complex; however, the proteolysis studies suggest that there are common structural features in their ligand binding sites. 相似文献
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Control of transcription activation by steroid hormone receptors. 总被引:11,自引:0,他引:11
H Gronemeyer 《FASEB journal》1992,6(8):2524-2529
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Ligand-dependent cleavage of the P75 neurotrophin receptor is necessary for NRIF nuclear translocation and apoptosis in sympathetic neurons 总被引:3,自引:0,他引:3
The p75 neurotrophin receptor regulates neuronal survival, promoting it in some contexts yet activating apoptosis in others. The mechanism by which the receptor elicits these differential effects is poorly understood. Here, we demonstrate that p75 is cleaved by gamma-secretase in sympathetic neurons, specifically in response to proapoptotic ligands. This cleavage resulted in ubiquitination and subsequent nuclear translocation of NRIF, a DNA binding protein essential for p75-mediated apoptosis. Inhibition of gamma-secretase or expression of a mutant p75 resistant to this protease prevented receptor proteolysis, blocked NRIF nuclear entry, and prevented apoptosis. In contrast, overexpression of the p75 ICD resulted in NRIF nuclear accumulation and apoptosis. The receptor proteolysis and NRIF nuclear localization were also observed in vivo during naturally occurring cell death in the superior cervical ganglia. These results indicate that p75-mediated apoptosis requires gamma-secretase dependent release of its ICD, which facilitates nuclear translocation of NRIF. 相似文献