Proteomic identification of glucocorticoid receptor interacting proteins. Hedman, E., Widen, C., Asadi, A., Dinnetz, I., Schroder, W. P., Gustafsson, J., & Wikstrom, A. Proteomics, 6(10):3114–3126, May, 2006. Place: Hoboken Publisher: Wiley WOS:000238010400018
doi  abstract   bibtex   
The glucocorticoid receptor (GR) acts as a ligand dependent transcription factor but can also cross talk with other signaling pathways via protein-protein interactions. In this paper we describe methods to study novel cytosolic GR interacting proteins, using mAb based immunoaffinity chromatography of GR from rat liver cytosol. Co-purifying proteins were identified by 2-DE in combination with MALDI-TOF-MS. Non-liganded/non-activated and in vitro liganded/ activated GR, respectively, co-purifies with specific sets of proteins. Of these 34 were conclusively identified, seven have previously been reported to be part of the GR-complex, revealing 27 new possible interacting candidates for the GR-complex. Of the novel GR interacting proteins the major vault protein, TATA binding interacting protein 49a and glycoprotein PP63 were of special interest. Furthermore, using 2-D DIGE we show that the set of proteins interacting with nonliganded GR is distinctly different in protein amount compared to the proteins found with liganded/activated GR. This suggests the presence of different GR complexes in the cell, which was further substantiated by the finding of several separate GR native protein complexes, "GR-receptosomes", using blue native gel electrophoresis. Our findings suggest the existence of several new mechanisms for GR signaling and regulation.
@article{hedman_proteomic_2006,
	title = {Proteomic identification of glucocorticoid receptor interacting proteins},
	volume = {6},
	issn = {1615-9853},
	doi = {10.1002/pmic.200500266},
	abstract = {The glucocorticoid receptor (GR) acts as a ligand dependent transcription factor but can also cross talk with other signaling pathways via protein-protein interactions. In this paper we describe methods to study novel cytosolic GR interacting proteins, using mAb based immunoaffinity chromatography of GR from rat liver cytosol. Co-purifying proteins were identified by 2-DE in combination with MALDI-TOF-MS. Non-liganded/non-activated and in vitro liganded/ activated GR, respectively, co-purifies with specific sets of proteins. Of these 34 were conclusively identified, seven have previously been reported to be part of the GR-complex, revealing 27 new possible interacting candidates for the GR-complex. Of the novel GR interacting proteins the major vault protein, TATA binding interacting protein 49a and glycoprotein PP63 were of special interest. Furthermore, using 2-D DIGE we show that the set of proteins interacting with nonliganded GR is distinctly different in protein amount compared to the proteins found with liganded/activated GR. This suggests the presence of different GR complexes in the cell, which was further substantiated by the finding of several separate GR native protein complexes, "GR-receptosomes", using blue native gel electrophoresis. Our findings suggest the existence of several new mechanisms for GR signaling and regulation.},
	language = {English},
	number = {10},
	journal = {Proteomics},
	author = {Hedman, Erik and Widen, Christina and Asadi, Abolfazl and Dinnetz, Ingrid and Schroder, Wolfgang P. and Gustafsson, Jan-ke and Wikstrom, Ann-Charlotte},
	month = may,
	year = {2006},
	note = {Place: Hoboken
Publisher: Wiley
WOS:000238010400018},
	keywords = {2-D blue native-PAGE, 2-d dige, 2-de, Animals, Antibodies, Monoclonal, Cell Line, Tumor, Chromatography, Affinity, Cytosol, Electrophoresis, Gel, Two-Dimensional, Electrophoresis, Polyacrylamide Gel, Immunoblotting, Ligands, Liver, Protein Interaction Mapping, Proteome, Rats, Receptors, Glucocorticoid, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, dna-binding, electrophoresis, glucocorticoid receptor, heat-shock-protein, kappa-b, living cells, localization, maldi-tof, mechanism, polyacrylamide-gels, rat, tyrosine kinase},
	pages = {3114--3126},
}

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