Phosphorylation of immunopurified rat liver glucocorticoid receptor by the catalytic subunit of cAMP-dependent protein kinase
Abstract
We have examined phosphorylation of the rat liver glucocorticoid receptor (GR) and GR-associated protein kinase (PK) activity in the immunopurified receptor preparations. Affinity labeling of hepatic cytosol with [3H]dexamethasone 21-mesylate showed a covalent association of the steroid with a 94 kDa protein. GR was immunopurified with antireceptor monoclonal antibody BuGR2 (Gametchu & Harrison, Endocrinology 114: 274-279, 1984) to near homogeneity. A 23 degrees C incubation of the immunoprecipitated protein A-Sepharose adsorbed GR with [gamma-32P]ATP,Mg2+ and the catalytic subunit of cAMP-dependent PK (cAMP-PK) from bovine heart, led to an incorporation of radioactivity in the 94 kDa protein. Phosphorylation of GR was not evident in the absence of the added kinase. Of the radioinert nucleotides (ATP, GTP, UTP or CTP) tested, only ATP successfully competed with [gamma-32P]ATP demonstrating a nucleotide specific requirement for the phosphorylation of GR. Other divalent cations, such as Mn2+ or Ca2+, could not be substituted for Mg2+ during the phosphorylation reaction. Phosphorylation of GR was sensitive to the presence of the protein kinase inhibitor, H-8, an isoquinoline sulfonamide derivative. In addition, the incorporation of radioactivity into GR was both time- and temperature-dependent. The phosphorylation of GR by cAMP-PK was independent of the presence of hsp-90 and transformation state of the receptor. The results of this study demonstrate that GR is an effective subs…
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PKA phosphorylates GR independent of relevant chaperone proteins (e.g., hsp90).
“PKA has been shown to phosphorylate GR independent of the presence of relevant chaperone proteins (i.e., hsp90)”
From Cytokines and Glucocorticoid Receptor Signaling (2009) · cites this paper as [108]
The transformational state of GR and phosphorylation of GR by PKA can be inhibited by H-8, a PKA inhibitor.
“the transformational state of GR and phosphorylation of GR by PKA can be inhibited by H-8, a PKA inhibitor.108”
From Cytokines and Glucocorticoid Receptor Signaling (2009) · cites this paper as [108]
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Sorger, P. K., & Pelham, H. R. B. (1988) · Cell · doi:10.1016/s0092-8674(88)91219-6 · Cited by 832 · see citing works
Oikarinen, J., Hämäläinen, L., & Oikarinen, A. (1984) · Biochimica et Biophysica Acta (BBA) - General Subjects · doi:10.1016/0304-4165(84)90290-3 · Cited by 62 · see citing works
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Haroon, E., Raison, C. L., & Miller, A. H. (2012) · Neuropsychopharmacology · doi:10.1038/npp.2011.205 · Cited by 1,000 · see citing works
Pace, T. W. W., Hu, F., & Miller, A. H. (2007) · Brain, Behavior, and Immunity · doi:10.1016/j.bbi.2006.08.009 · Cited by 621 · see citing works
Cytokines and Glucocorticoid Receptor Signaling
Pace, T. W. W., & Miller, A. H. (2009) · Annals of the New York Academy of Sciences · doi:10.1111/j.1749-6632.2009.04984.x · Cited by 339 · see citing works
Beck, I. M. E., Vanden Berghe, W., Vermeulen, L., Yamamoto, K. R., Haegeman, G., & De Bosscher, K. (2009) · Endocrine Reviews · doi:10.1210/er.2009-0013 · Cited by 298 · see citing works
Doucas, V., Shi, Y., Miyamoto, S., West, A., Verma, I., & Evans, R. M. (2000) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.220413297 · Cited by 125 · see citing works
- Authors
- Haske, T., Nakao, M., & Moudgil, V. K.
- Affiliations
- Oakland University (US)
- Venue
- Molecular and Cellular Biochemistry
- Published
- 1994
- Volume
- 132
- Issue
- 2
- Pages
- 163-171
- ISSN
- 0300-8177 (ISSN-L)
- 1573-4919
- OpenAlex
- W2009959920
- Cited by
- 24
- References
- 2
- Citations by year
- 2023: 1 · 2022: 1 · 2017: 1 · 2016: 1 · 2015: 1 · 2013: 3 · 2012: 1
- FWCI
- 0.00
- Type
- journal article
- Language
- en
- Open access
- closed
- Licence
- www.springer.com/tdm (tdm)
- Locations
- Molecular and Cellular Biochemistry (publishedVersion)
- PubMed (publishedVersion)
- Funders
- National Institutes of Health
- National Institute of Diabetes and Digestive and Kidney Diseases (DK-20893, R01DK020893)
- Topics
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Estrogen and related hormone effects
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
- Keywords
- glucocorticoid receptor, receptor phosphorylation, protein kinase A, catalytic subunit, rat liver, immunoaffinity purification, ATP requirement, magnesium dependence, receptor kinase activity
- MeSH terms
- Animals, Binding Sites, Catalysis, Liver, Male, Phosphorylation, Precipitin Tests, Receptors, Glucocorticoid, Cyclic AMP-Dependent Protein Kinases, Rats
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