Recombinant human melatonin receptor MT1 isolated in mixed detergents shows pharmacology similar to that in mammalian cell membranes.

The human melatonin MT1 receptor-belonging to the large family of G protein-coupled receptors (GPCRs)-plays a key role in circadian rhythm regulation and is notably involved in sleep disorders and depression. Structural and functional information at the molecular level are highly desired for fine ch...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Christel Logez, Sylvie Berger, Céline Legros, Jean-Louis Banères, William Cohen, Philippe Delagrange, Olivier Nosjean, Jean A Boutin, Gilles Ferry, Frédéric Simonin, Renaud Wagner
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2014
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/961a2e8080f24c94a99bea2e9fb8eaad
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:961a2e8080f24c94a99bea2e9fb8eaad
record_format	dspace
spelling	oai:doaj.org-article:961a2e8080f24c94a99bea2e9fb8eaad2021-11-11T08:21:54ZRecombinant human melatonin receptor MT1 isolated in mixed detergents shows pharmacology similar to that in mammalian cell membranes.1932-620310.1371/journal.pone.0100616https://doaj.org/article/961a2e8080f24c94a99bea2e9fb8eaad2014-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24959712/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203The human melatonin MT1 receptor-belonging to the large family of G protein-coupled receptors (GPCRs)-plays a key role in circadian rhythm regulation and is notably involved in sleep disorders and depression. Structural and functional information at the molecular level are highly desired for fine characterization of this receptor; however, adequate techniques for isolating soluble MT1 material suitable for biochemical and biophysical studies remain lacking. Here we describe the evaluation of a panel of constructs and host systems for the production of recombinant human MT1 receptors, and the screening of different conditions for their solubilization and purification. Our findings resulted in the establishment of an original strategy using a mixture of Fos14 and CHAPS detergents to extract and purify a recombinant human MT1 from Pichia pastoris membranes. This procedure enabled the recovery of relatively pure, monomeric and ligand-binding active MT1 receptor in the near-milligram range. A comparative study based on extensive ligand-binding characterization highlighted a very close correlation between the pharmacological profiles of MT1 purified from yeast and the same receptor present in mammalian cell membranes. The high quality of the purified MT1 was further confirmed by its ability to activate its cognate Gαi protein partner when reconstituted in lipid discs, thus opening novel paths to investigate this receptor by biochemical and biophysical approaches.Christel LogezSylvie BergerCéline LegrosJean-Louis BanèresWilliam CohenPhilippe DelagrangeOlivier NosjeanJean A BoutinGilles FerryFrédéric SimoninRenaud WagnerPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 9, Iss 6, p e100616 (2014)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Christel Logez Sylvie Berger Céline Legros Jean-Louis Banères William Cohen Philippe Delagrange Olivier Nosjean Jean A Boutin Gilles Ferry Frédéric Simonin Renaud Wagner Recombinant human melatonin receptor MT1 isolated in mixed detergents shows pharmacology similar to that in mammalian cell membranes.
description	The human melatonin MT1 receptor-belonging to the large family of G protein-coupled receptors (GPCRs)-plays a key role in circadian rhythm regulation and is notably involved in sleep disorders and depression. Structural and functional information at the molecular level are highly desired for fine characterization of this receptor; however, adequate techniques for isolating soluble MT1 material suitable for biochemical and biophysical studies remain lacking. Here we describe the evaluation of a panel of constructs and host systems for the production of recombinant human MT1 receptors, and the screening of different conditions for their solubilization and purification. Our findings resulted in the establishment of an original strategy using a mixture of Fos14 and CHAPS detergents to extract and purify a recombinant human MT1 from Pichia pastoris membranes. This procedure enabled the recovery of relatively pure, monomeric and ligand-binding active MT1 receptor in the near-milligram range. A comparative study based on extensive ligand-binding characterization highlighted a very close correlation between the pharmacological profiles of MT1 purified from yeast and the same receptor present in mammalian cell membranes. The high quality of the purified MT1 was further confirmed by its ability to activate its cognate Gαi protein partner when reconstituted in lipid discs, thus opening novel paths to investigate this receptor by biochemical and biophysical approaches.
format	article
author	Christel Logez Sylvie Berger Céline Legros Jean-Louis Banères William Cohen Philippe Delagrange Olivier Nosjean Jean A Boutin Gilles Ferry Frédéric Simonin Renaud Wagner
author_facet	Christel Logez Sylvie Berger Céline Legros Jean-Louis Banères William Cohen Philippe Delagrange Olivier Nosjean Jean A Boutin Gilles Ferry Frédéric Simonin Renaud Wagner
author_sort	Christel Logez
title	Recombinant human melatonin receptor MT1 isolated in mixed detergents shows pharmacology similar to that in mammalian cell membranes.
title_short	Recombinant human melatonin receptor MT1 isolated in mixed detergents shows pharmacology similar to that in mammalian cell membranes.
title_full	Recombinant human melatonin receptor MT1 isolated in mixed detergents shows pharmacology similar to that in mammalian cell membranes.
title_fullStr	Recombinant human melatonin receptor MT1 isolated in mixed detergents shows pharmacology similar to that in mammalian cell membranes.
title_full_unstemmed	Recombinant human melatonin receptor MT1 isolated in mixed detergents shows pharmacology similar to that in mammalian cell membranes.
title_sort	recombinant human melatonin receptor mt1 isolated in mixed detergents shows pharmacology similar to that in mammalian cell membranes.
publisher	Public Library of Science (PLoS)
publishDate	2014
url	https://doaj.org/article/961a2e8080f24c94a99bea2e9fb8eaad
work_keys_str_mv	AT christellogez recombinanthumanmelatoninreceptormt1isolatedinmixeddetergentsshowspharmacologysimilartothatinmammaliancellmembranes AT sylvieberger recombinanthumanmelatoninreceptormt1isolatedinmixeddetergentsshowspharmacologysimilartothatinmammaliancellmembranes AT celinelegros recombinanthumanmelatoninreceptormt1isolatedinmixeddetergentsshowspharmacologysimilartothatinmammaliancellmembranes AT jeanlouisbaneres recombinanthumanmelatoninreceptormt1isolatedinmixeddetergentsshowspharmacologysimilartothatinmammaliancellmembranes AT williamcohen recombinanthumanmelatoninreceptormt1isolatedinmixeddetergentsshowspharmacologysimilartothatinmammaliancellmembranes AT philippedelagrange recombinanthumanmelatoninreceptormt1isolatedinmixeddetergentsshowspharmacologysimilartothatinmammaliancellmembranes AT oliviernosjean recombinanthumanmelatoninreceptormt1isolatedinmixeddetergentsshowspharmacologysimilartothatinmammaliancellmembranes AT jeanaboutin recombinanthumanmelatoninreceptormt1isolatedinmixeddetergentsshowspharmacologysimilartothatinmammaliancellmembranes AT gillesferry recombinanthumanmelatoninreceptormt1isolatedinmixeddetergentsshowspharmacologysimilartothatinmammaliancellmembranes AT fredericsimonin recombinanthumanmelatoninreceptormt1isolatedinmixeddetergentsshowspharmacologysimilartothatinmammaliancellmembranes AT renaudwagner recombinanthumanmelatoninreceptormt1isolatedinmixeddetergentsshowspharmacologysimilartothatinmammaliancellmembranes
_version_	1718439332688166912

Recombinant human melatonin receptor MT1 isolated in mixed detergents shows pharmacology similar to that in mammalian cell membranes.

Ejemplares similares