Stability study on renal type I mineralocorticoid receptor
The purpose of this work is to review stability and activation properties of type 1 receptor, in order to explain the reasons for its extreme in vitro instability. We demonstrate that the treatment of rat kidney cytosol with H2O2 prevents aldosterone binding, DNA/steroid- receptor complex interactio...
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| Acceso en línea: | http://hdl.handle.net/20.500.12110/paper_00243205_v59_n7_p511_Galigniana |
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todo:paper_00243205_v59_n7_p511_Galigniana2023-10-03T14:34:40Z Stability study on renal type I mineralocorticoid receptor Galigniana, M.D. mineralocorticoid receptor oxidation stability thiol groups 5,5' dithiobis(2 nitrobenzoic acid) aldosterone DNA edetic acid egtazic acid hydrogen peroxide iron mercaptoethanol mineralocorticoid receptor n ethylmaleimide steroid thiol group animal cell article cell free system cytosol DNA binding high temperature procedures kidney nonhuman oxidation rat receptor binding thermostability Animalia The purpose of this work is to review stability and activation properties of type 1 receptor, in order to explain the reasons for its extreme in vitro instability. We demonstrate that the treatment of rat kidney cytosol with H2O2 prevents aldosterone binding, DNA/steroid- receptor complex interactions, and prevents the receptor thermal inactivation. In contrast, exogenous sulfhydryl reducing reagents are necessary to insure maximum binding of mineralocorticoid receptor and DNA/steroid-receptor interaction. However, the presence of β- mercaptoethanol in thermal induced incubations reverts the H2O2 protection. We also demonstrate that contaminations with free or sequestered iron are harmful for both, receptor binding capacity (in a reversible form) and for hormone-receptor/DNA binding properties (in a partially reversible form). We propose a sulfhydryl oxidative mechanism for type I mineralocorticoid receptor inactivation in which iron contaminants might accelerate this process by oxidative catalysis. We also demonstrate that when thiol groups are blocked by specific reagents such as N-ethyl-maleimide or dithionitrobenzoic acid, type I sites loose binding capacity, but the protein is protected from oxidation as well as inactivation. JOUR info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by/2.5/ar http://hdl.handle.net/20.500.12110/paper_00243205_v59_n7_p511_Galigniana |
| institution |
Universidad de Buenos Aires |
| institution_str |
I-28 |
| repository_str |
R-134 |
| collection |
Biblioteca Digital - Facultad de Ciencias Exactas y Naturales (UBA) |
| topic |
mineralocorticoid receptor oxidation stability thiol groups 5,5' dithiobis(2 nitrobenzoic acid) aldosterone DNA edetic acid egtazic acid hydrogen peroxide iron mercaptoethanol mineralocorticoid receptor n ethylmaleimide steroid thiol group animal cell article cell free system cytosol DNA binding high temperature procedures kidney nonhuman oxidation rat receptor binding thermostability Animalia |
| spellingShingle |
mineralocorticoid receptor oxidation stability thiol groups 5,5' dithiobis(2 nitrobenzoic acid) aldosterone DNA edetic acid egtazic acid hydrogen peroxide iron mercaptoethanol mineralocorticoid receptor n ethylmaleimide steroid thiol group animal cell article cell free system cytosol DNA binding high temperature procedures kidney nonhuman oxidation rat receptor binding thermostability Animalia Galigniana, M.D. Stability study on renal type I mineralocorticoid receptor |
| topic_facet |
mineralocorticoid receptor oxidation stability thiol groups 5,5' dithiobis(2 nitrobenzoic acid) aldosterone DNA edetic acid egtazic acid hydrogen peroxide iron mercaptoethanol mineralocorticoid receptor n ethylmaleimide steroid thiol group animal cell article cell free system cytosol DNA binding high temperature procedures kidney nonhuman oxidation rat receptor binding thermostability Animalia |
| description |
The purpose of this work is to review stability and activation properties of type 1 receptor, in order to explain the reasons for its extreme in vitro instability. We demonstrate that the treatment of rat kidney cytosol with H2O2 prevents aldosterone binding, DNA/steroid- receptor complex interactions, and prevents the receptor thermal inactivation. In contrast, exogenous sulfhydryl reducing reagents are necessary to insure maximum binding of mineralocorticoid receptor and DNA/steroid-receptor interaction. However, the presence of β- mercaptoethanol in thermal induced incubations reverts the H2O2 protection. We also demonstrate that contaminations with free or sequestered iron are harmful for both, receptor binding capacity (in a reversible form) and for hormone-receptor/DNA binding properties (in a partially reversible form). We propose a sulfhydryl oxidative mechanism for type I mineralocorticoid receptor inactivation in which iron contaminants might accelerate this process by oxidative catalysis. We also demonstrate that when thiol groups are blocked by specific reagents such as N-ethyl-maleimide or dithionitrobenzoic acid, type I sites loose binding capacity, but the protein is protected from oxidation as well as inactivation. |
| format |
JOUR |
| author |
Galigniana, M.D. |
| author_facet |
Galigniana, M.D. |
| author_sort |
Galigniana, M.D. |
| title |
Stability study on renal type I mineralocorticoid receptor |
| title_short |
Stability study on renal type I mineralocorticoid receptor |
| title_full |
Stability study on renal type I mineralocorticoid receptor |
| title_fullStr |
Stability study on renal type I mineralocorticoid receptor |
| title_full_unstemmed |
Stability study on renal type I mineralocorticoid receptor |
| title_sort |
stability study on renal type i mineralocorticoid receptor |
| url |
http://hdl.handle.net/20.500.12110/paper_00243205_v59_n7_p511_Galigniana |
| work_keys_str_mv |
AT galignianamd stabilitystudyonrenaltypeimineralocorticoidreceptor |
| _version_ |
1807322392225120256 |