Modification of an essential amino group in the mineralocorticoid receptor evidences a differential conformational change of the receptor protein upon binding of antagonists, natural agonists and the synthetic agonist 11,19-oxidoprogesterone
The alkylation of amino groups of the mineralocorticoid receptor (MR) with pyridoxal 5′-phosphate or 2,4,6-trinitrobenzenesulphonate (TNBS) under controlled conditions modifies only one lysyl residue, which accounts for a 70% inhibition of steroid binding capacity. The Kd of aldosterone for MR is no...
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| Formato: | Artículo publishedVersion |
| Lenguaje: | Inglés |
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2002
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| Acceso en línea: | http://hdl.handle.net/20.500.12110/paper_01674889_v1589_n1_p31_PiwienPilipuk |
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paperaa:paper_01674889_v1589_n1_p31_PiwienPilipuk2023-06-12T16:46:57Z Modification of an essential amino group in the mineralocorticoid receptor evidences a differential conformational change of the receptor protein upon binding of antagonists, natural agonists and the synthetic agonist 11,19-oxidoprogesterone Biochim. Biophys. Acta Mol. Cell Res. 2002;1589(1):31-48 Piwien-Pilipuk, G. Kanelakis, K.C. Ghini, A.A. Lantos, C.P. Litwack, G. Burton, G. Galigniana, M.D. 2,4,6-Trinitrobenzenesulfonate Aldosterone hsp90 heterocomplex Pyridoxal 5′-phosphate Sodium retention 11,19 oxidoprogesterone 21 deoxypregnanesteroid 11,19 oxidoprogesterone 6,19 oxidoprogesterone aldosterone alkylating agent amino acid chymotrypsin A corticosterone deoxycorticosterone heat shock protein 90 ligand lysine mineralocorticoid antagonist mineralocorticoid receptor progesterone derivative pyridoxal 5 phosphate receptor protein spironolactone testosterone trinitrobenzenesulfonic acid unclassified drug alkylation animal tissue article binding affinity binding site chemical modification conformational transition controlled study inhibition kinetics ligand binding male nonhuman priority journal protein degradation rat receptor binding steroid binding Aldosterone Alkylating Agents Amines Animals Binding Sites Chymotrypsin Cytosol HSP90 Heat-Shock Proteins Hydrogen-Ion Concentration Kidney Male Progesterone Protein Conformation Pyridoxal Phosphate Rats Rats, Sprague-Dawley Receptors, Mineralocorticoid Time Factors Trinitrobenzenesulfonic Acid Tritium The alkylation of amino groups of the mineralocorticoid receptor (MR) with pyridoxal 5′-phosphate or 2,4,6-trinitrobenzenesulphonate (TNBS) under controlled conditions modifies only one lysyl residue, which accounts for a 70% inhibition of steroid binding capacity. The Kd of aldosterone for MR is not affected by the treatment, but the total number of binding sites is greatly decreased. The modified receptor is capable of dynamically conserving its association with the hsp90-based heterocomplex. Importantly, the binding of natural agonists protects the hormone binding capacity of the MR from the inactivating action of alkylating agents. In contrast, antagonistic steroids are totally incapable of providing such protection. Like the antagonistic ligands, and despite its potent mineralocorticoid biological effect, the sole MR specific synthetic agonist known to date, 11,19-oxidoprogesterone (11-OP), shows no protective effect upon treatment of the MR with pyridoxal 5′-phosphate or TNBS. Limited digestion of the MR with α-chymotrypsin generates a 34 kDa fragment, which becomes totally resistant to digestion upon binding of natural agonists, but not upon binding of antagonists. Interestingly, the synthetic 21-deoxypregnanesteroid 11-OP exhibits an intermediate pattern of proteolytic degradation, suggesting that the conformational change generated in the MR is not equivalent to that induced by antagonists or natural agonists. We conclude that in the first steps of activation, the MR changes its conformation upon binding of the ligand. However, the nature of this conformational change depends on the nature of the ligand. The experimental evidence shown in this work suggests that a single lysyl group can determine the hormone specificity of the MR. © 2002 Elsevier Science B.V. All rights reserved. Fil:Ghini, A.A. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. Fil:Burton, G. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina. 2002 info:eu-repo/semantics/article info:ar-repo/semantics/artículo info:eu-repo/semantics/publishedVersion application/pdf eng info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_01674889_v1589_n1_p31_PiwienPilipuk |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| language |
Inglés |
| orig_language_str_mv |
eng |
| topic |
2,4,6-Trinitrobenzenesulfonate Aldosterone hsp90 heterocomplex Pyridoxal 5′-phosphate Sodium retention 11,19 oxidoprogesterone 21 deoxypregnanesteroid 11,19 oxidoprogesterone 6,19 oxidoprogesterone aldosterone alkylating agent amino acid chymotrypsin A corticosterone deoxycorticosterone heat shock protein 90 ligand lysine mineralocorticoid antagonist mineralocorticoid receptor progesterone derivative pyridoxal 5 phosphate receptor protein spironolactone testosterone trinitrobenzenesulfonic acid unclassified drug alkylation animal tissue article binding affinity binding site chemical modification conformational transition controlled study inhibition kinetics ligand binding male nonhuman priority journal protein degradation rat receptor binding steroid binding Aldosterone Alkylating Agents Amines Animals Binding Sites Chymotrypsin Cytosol HSP90 Heat-Shock Proteins Hydrogen-Ion Concentration Kidney Male Progesterone Protein Conformation Pyridoxal Phosphate Rats Rats, Sprague-Dawley Receptors, Mineralocorticoid Time Factors Trinitrobenzenesulfonic Acid Tritium |
| spellingShingle |
2,4,6-Trinitrobenzenesulfonate Aldosterone hsp90 heterocomplex Pyridoxal 5′-phosphate Sodium retention 11,19 oxidoprogesterone 21 deoxypregnanesteroid 11,19 oxidoprogesterone 6,19 oxidoprogesterone aldosterone alkylating agent amino acid chymotrypsin A corticosterone deoxycorticosterone heat shock protein 90 ligand lysine mineralocorticoid antagonist mineralocorticoid receptor progesterone derivative pyridoxal 5 phosphate receptor protein spironolactone testosterone trinitrobenzenesulfonic acid unclassified drug alkylation animal tissue article binding affinity binding site chemical modification conformational transition controlled study inhibition kinetics ligand binding male nonhuman priority journal protein degradation rat receptor binding steroid binding Aldosterone Alkylating Agents Amines Animals Binding Sites Chymotrypsin Cytosol HSP90 Heat-Shock Proteins Hydrogen-Ion Concentration Kidney Male Progesterone Protein Conformation Pyridoxal Phosphate Rats Rats, Sprague-Dawley Receptors, Mineralocorticoid Time Factors Trinitrobenzenesulfonic Acid Tritium Piwien-Pilipuk, G. Kanelakis, K.C. Ghini, A.A. Lantos, C.P. Litwack, G. Burton, G. Galigniana, M.D. Modification of an essential amino group in the mineralocorticoid receptor evidences a differential conformational change of the receptor protein upon binding of antagonists, natural agonists and the synthetic agonist 11,19-oxidoprogesterone |
| topic_facet |
2,4,6-Trinitrobenzenesulfonate Aldosterone hsp90 heterocomplex Pyridoxal 5′-phosphate Sodium retention 11,19 oxidoprogesterone 21 deoxypregnanesteroid 11,19 oxidoprogesterone 6,19 oxidoprogesterone aldosterone alkylating agent amino acid chymotrypsin A corticosterone deoxycorticosterone heat shock protein 90 ligand lysine mineralocorticoid antagonist mineralocorticoid receptor progesterone derivative pyridoxal 5 phosphate receptor protein spironolactone testosterone trinitrobenzenesulfonic acid unclassified drug alkylation animal tissue article binding affinity binding site chemical modification conformational transition controlled study inhibition kinetics ligand binding male nonhuman priority journal protein degradation rat receptor binding steroid binding Aldosterone Alkylating Agents Amines Animals Binding Sites Chymotrypsin Cytosol HSP90 Heat-Shock Proteins Hydrogen-Ion Concentration Kidney Male Progesterone Protein Conformation Pyridoxal Phosphate Rats Rats, Sprague-Dawley Receptors, Mineralocorticoid Time Factors Trinitrobenzenesulfonic Acid Tritium |
| description |
The alkylation of amino groups of the mineralocorticoid receptor (MR) with pyridoxal 5′-phosphate or 2,4,6-trinitrobenzenesulphonate (TNBS) under controlled conditions modifies only one lysyl residue, which accounts for a 70% inhibition of steroid binding capacity. The Kd of aldosterone for MR is not affected by the treatment, but the total number of binding sites is greatly decreased. The modified receptor is capable of dynamically conserving its association with the hsp90-based heterocomplex. Importantly, the binding of natural agonists protects the hormone binding capacity of the MR from the inactivating action of alkylating agents. In contrast, antagonistic steroids are totally incapable of providing such protection. Like the antagonistic ligands, and despite its potent mineralocorticoid biological effect, the sole MR specific synthetic agonist known to date, 11,19-oxidoprogesterone (11-OP), shows no protective effect upon treatment of the MR with pyridoxal 5′-phosphate or TNBS. Limited digestion of the MR with α-chymotrypsin generates a 34 kDa fragment, which becomes totally resistant to digestion upon binding of natural agonists, but not upon binding of antagonists. Interestingly, the synthetic 21-deoxypregnanesteroid 11-OP exhibits an intermediate pattern of proteolytic degradation, suggesting that the conformational change generated in the MR is not equivalent to that induced by antagonists or natural agonists. We conclude that in the first steps of activation, the MR changes its conformation upon binding of the ligand. However, the nature of this conformational change depends on the nature of the ligand. The experimental evidence shown in this work suggests that a single lysyl group can determine the hormone specificity of the MR. © 2002 Elsevier Science B.V. All rights reserved. |
| format |
Artículo Artículo publishedVersion |
| author |
Piwien-Pilipuk, G. Kanelakis, K.C. Ghini, A.A. Lantos, C.P. Litwack, G. Burton, G. Galigniana, M.D. |
| author_facet |
Piwien-Pilipuk, G. Kanelakis, K.C. Ghini, A.A. Lantos, C.P. Litwack, G. Burton, G. Galigniana, M.D. |
| author_sort |
Piwien-Pilipuk, G. |
| title |
Modification of an essential amino group in the mineralocorticoid receptor evidences a differential conformational change of the receptor protein upon binding of antagonists, natural agonists and the synthetic agonist 11,19-oxidoprogesterone |
| title_short |
Modification of an essential amino group in the mineralocorticoid receptor evidences a differential conformational change of the receptor protein upon binding of antagonists, natural agonists and the synthetic agonist 11,19-oxidoprogesterone |
| title_full |
Modification of an essential amino group in the mineralocorticoid receptor evidences a differential conformational change of the receptor protein upon binding of antagonists, natural agonists and the synthetic agonist 11,19-oxidoprogesterone |
| title_fullStr |
Modification of an essential amino group in the mineralocorticoid receptor evidences a differential conformational change of the receptor protein upon binding of antagonists, natural agonists and the synthetic agonist 11,19-oxidoprogesterone |
| title_full_unstemmed |
Modification of an essential amino group in the mineralocorticoid receptor evidences a differential conformational change of the receptor protein upon binding of antagonists, natural agonists and the synthetic agonist 11,19-oxidoprogesterone |
| title_sort |
modification of an essential amino group in the mineralocorticoid receptor evidences a differential conformational change of the receptor protein upon binding of antagonists, natural agonists and the synthetic agonist 11,19-oxidoprogesterone |
| publishDate |
2002 |
| url |
http://hdl.handle.net/20.500.12110/paper_01674889_v1589_n1_p31_PiwienPilipuk |
| work_keys_str_mv |
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