Identification of threonine 422 in transmembrane domain αM4 of the nicotinic acetylcholine receptor as a possible site of interaction with hydrocortisone
- Autores
- Garbus, Ingrid; Roccamo, Ana Maria; Barrantes, Francisco Jose
- Año de publicación
- 2002
- Idioma
- inglés
- Tipo de recurso
- artículo
- Estado
- versión publicada
- Descripción
- The modulatory effects exerted by the glucocorticoid hydrocortisone (HC) on the nicotinic acetylcholine receptor (AChR) were studied in mutants of the α subunit M4 transmembrane region. Based on the photoaffinity labeling of αM4 412 with the steroid promegestone this position was mutated to different residues to explore the properties of side-chain volume, hydrophobicity, and charge on AChR-steroid interactions. All mutants showed channel kinetics indistinguishable from those of the wild-type AChR, both in the absence and presence of HC (200 and 400 μM), in single-channel recordings at different acetylcholine (ACh) concentrations. An alanine-substituted quadruple mutant of four putative lipid-exposed residues in αM4 (L411, M415, C418 and T422) exhibited less inhibition by HC than that observed in wild-type AChR. When we dissected the quadruple mutant into four individual alanine-substituted receptors, we found that the T422 mutant AChR behaved like the quadruple mutant, whereas the other three were indistinguishable from the wild-type. We conclude that T422, a residue close to the extracellular-facing membrane hemilayer in αM4, has direct bearing on the changes in HC sensitivity and propose its involvement in the steroid-AChR interaction site.
Fil: Garbus, Ingrid. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Unesco; Argentina
Fil: Roccamo, Ana Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Unesco; Argentina
Fil: Barrantes, Francisco Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Unesco; Argentina - Materia
-
Cholinergic Receptor
Glucocorticoid
Mutations
Patch-Clamp
Steroid - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
- Repositorio
.jpg)
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/53088
Ver los metadatos del registro completo
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Identification of threonine 422 in transmembrane domain αM4 of the nicotinic acetylcholine receptor as a possible site of interaction with hydrocortisoneGarbus, IngridRoccamo, Ana MariaBarrantes, Francisco JoseCholinergic ReceptorGlucocorticoidMutationsPatch-ClampSteroidhttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The modulatory effects exerted by the glucocorticoid hydrocortisone (HC) on the nicotinic acetylcholine receptor (AChR) were studied in mutants of the α subunit M4 transmembrane region. Based on the photoaffinity labeling of αM4 412 with the steroid promegestone this position was mutated to different residues to explore the properties of side-chain volume, hydrophobicity, and charge on AChR-steroid interactions. All mutants showed channel kinetics indistinguishable from those of the wild-type AChR, both in the absence and presence of HC (200 and 400 μM), in single-channel recordings at different acetylcholine (ACh) concentrations. An alanine-substituted quadruple mutant of four putative lipid-exposed residues in αM4 (L411, M415, C418 and T422) exhibited less inhibition by HC than that observed in wild-type AChR. When we dissected the quadruple mutant into four individual alanine-substituted receptors, we found that the T422 mutant AChR behaved like the quadruple mutant, whereas the other three were indistinguishable from the wild-type. We conclude that T422, a residue close to the extracellular-facing membrane hemilayer in αM4, has direct bearing on the changes in HC sensitivity and propose its involvement in the steroid-AChR interaction site.Fil: Garbus, Ingrid. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Unesco; ArgentinaFil: Roccamo, Ana Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Unesco; ArgentinaFil: Barrantes, Francisco Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Unesco; ArgentinaPergamon-Elsevier Science Ltd2002-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/53088Garbus, Ingrid; Roccamo, Ana Maria; Barrantes, Francisco Jose; Identification of threonine 422 in transmembrane domain αM4 of the nicotinic acetylcholine receptor as a possible site of interaction with hydrocortisone; Pergamon-Elsevier Science Ltd; Neuropharmacology; 43; 1; 7-2002; 65-730028-3908CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0028390802000680info:eu-repo/semantics/altIdentifier/doi/10.1016/S0028-3908(02)00068-0info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-22T11:07:15Zoai:ri.conicet.gov.ar:11336/53088instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-10-22 11:07:15.601CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
| dc.title.none.fl_str_mv |
Identification of threonine 422 in transmembrane domain αM4 of the nicotinic acetylcholine receptor as a possible site of interaction with hydrocortisone |
| title |
Identification of threonine 422 in transmembrane domain αM4 of the nicotinic acetylcholine receptor as a possible site of interaction with hydrocortisone |
| spellingShingle |
Identification of threonine 422 in transmembrane domain αM4 of the nicotinic acetylcholine receptor as a possible site of interaction with hydrocortisone Garbus, Ingrid Cholinergic Receptor Glucocorticoid Mutations Patch-Clamp Steroid |
| title_short |
Identification of threonine 422 in transmembrane domain αM4 of the nicotinic acetylcholine receptor as a possible site of interaction with hydrocortisone |
| title_full |
Identification of threonine 422 in transmembrane domain αM4 of the nicotinic acetylcholine receptor as a possible site of interaction with hydrocortisone |
| title_fullStr |
Identification of threonine 422 in transmembrane domain αM4 of the nicotinic acetylcholine receptor as a possible site of interaction with hydrocortisone |
| title_full_unstemmed |
Identification of threonine 422 in transmembrane domain αM4 of the nicotinic acetylcholine receptor as a possible site of interaction with hydrocortisone |
| title_sort |
Identification of threonine 422 in transmembrane domain αM4 of the nicotinic acetylcholine receptor as a possible site of interaction with hydrocortisone |
| dc.creator.none.fl_str_mv |
Garbus, Ingrid Roccamo, Ana Maria Barrantes, Francisco Jose |
| author |
Garbus, Ingrid |
| author_facet |
Garbus, Ingrid Roccamo, Ana Maria Barrantes, Francisco Jose |
| author_role |
author |
| author2 |
Roccamo, Ana Maria Barrantes, Francisco Jose |
| author2_role |
author author |
| dc.subject.none.fl_str_mv |
Cholinergic Receptor Glucocorticoid Mutations Patch-Clamp Steroid |
| topic |
Cholinergic Receptor Glucocorticoid Mutations Patch-Clamp Steroid |
| purl_subject.fl_str_mv |
https://purl.org/becyt/ford/1.6 https://purl.org/becyt/ford/1 |
| dc.description.none.fl_txt_mv |
The modulatory effects exerted by the glucocorticoid hydrocortisone (HC) on the nicotinic acetylcholine receptor (AChR) were studied in mutants of the α subunit M4 transmembrane region. Based on the photoaffinity labeling of αM4 412 with the steroid promegestone this position was mutated to different residues to explore the properties of side-chain volume, hydrophobicity, and charge on AChR-steroid interactions. All mutants showed channel kinetics indistinguishable from those of the wild-type AChR, both in the absence and presence of HC (200 and 400 μM), in single-channel recordings at different acetylcholine (ACh) concentrations. An alanine-substituted quadruple mutant of four putative lipid-exposed residues in αM4 (L411, M415, C418 and T422) exhibited less inhibition by HC than that observed in wild-type AChR. When we dissected the quadruple mutant into four individual alanine-substituted receptors, we found that the T422 mutant AChR behaved like the quadruple mutant, whereas the other three were indistinguishable from the wild-type. We conclude that T422, a residue close to the extracellular-facing membrane hemilayer in αM4, has direct bearing on the changes in HC sensitivity and propose its involvement in the steroid-AChR interaction site. Fil: Garbus, Ingrid. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Unesco; Argentina Fil: Roccamo, Ana Maria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Unesco; Argentina Fil: Barrantes, Francisco Jose. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Bahía Blanca. Instituto de Investigaciones Bioquímicas de Bahía Blanca. Universidad Nacional del Sur. Instituto de Investigaciones Bioquímicas de Bahía Blanca; Argentina. Unesco; Argentina |
| description |
The modulatory effects exerted by the glucocorticoid hydrocortisone (HC) on the nicotinic acetylcholine receptor (AChR) were studied in mutants of the α subunit M4 transmembrane region. Based on the photoaffinity labeling of αM4 412 with the steroid promegestone this position was mutated to different residues to explore the properties of side-chain volume, hydrophobicity, and charge on AChR-steroid interactions. All mutants showed channel kinetics indistinguishable from those of the wild-type AChR, both in the absence and presence of HC (200 and 400 μM), in single-channel recordings at different acetylcholine (ACh) concentrations. An alanine-substituted quadruple mutant of four putative lipid-exposed residues in αM4 (L411, M415, C418 and T422) exhibited less inhibition by HC than that observed in wild-type AChR. When we dissected the quadruple mutant into four individual alanine-substituted receptors, we found that the T422 mutant AChR behaved like the quadruple mutant, whereas the other three were indistinguishable from the wild-type. We conclude that T422, a residue close to the extracellular-facing membrane hemilayer in αM4, has direct bearing on the changes in HC sensitivity and propose its involvement in the steroid-AChR interaction site. |
| publishDate |
2002 |
| dc.date.none.fl_str_mv |
2002-07 |
| dc.type.none.fl_str_mv |
info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://purl.org/coar/resource_type/c_6501 info:ar-repo/semantics/articulo |
| format |
article |
| status_str |
publishedVersion |
| dc.identifier.none.fl_str_mv |
http://hdl.handle.net/11336/53088 Garbus, Ingrid; Roccamo, Ana Maria; Barrantes, Francisco Jose; Identification of threonine 422 in transmembrane domain αM4 of the nicotinic acetylcholine receptor as a possible site of interaction with hydrocortisone; Pergamon-Elsevier Science Ltd; Neuropharmacology; 43; 1; 7-2002; 65-73 0028-3908 CONICET Digital CONICET |
| url |
http://hdl.handle.net/11336/53088 |
| identifier_str_mv |
Garbus, Ingrid; Roccamo, Ana Maria; Barrantes, Francisco Jose; Identification of threonine 422 in transmembrane domain αM4 of the nicotinic acetylcholine receptor as a possible site of interaction with hydrocortisone; Pergamon-Elsevier Science Ltd; Neuropharmacology; 43; 1; 7-2002; 65-73 0028-3908 CONICET Digital CONICET |
| dc.language.none.fl_str_mv |
eng |
| language |
eng |
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info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/pii/S0028390802000680 info:eu-repo/semantics/altIdentifier/doi/10.1016/S0028-3908(02)00068-0 |
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info:eu-repo/semantics/openAccess https://creativecommons.org/licenses/by-nc-sa/2.5/ar/ |
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openAccess |
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https://creativecommons.org/licenses/by-nc-sa/2.5/ar/ |
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application/pdf application/pdf application/pdf |
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Pergamon-Elsevier Science Ltd |
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Pergamon-Elsevier Science Ltd |
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reponame:CONICET Digital (CONICET) instname:Consejo Nacional de Investigaciones Científicas y Técnicas |
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Consejo Nacional de Investigaciones Científicas y Técnicas |
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CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas |
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dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar |
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