Haloferax volcanii, as a Novel Tool for Producing Mammalian Olfactory Receptors Embedded in Archaeal Lipid Bilayer

The aim of this study was to explore the possibility of using an archaeal microorganism as a host system for expressing mammalian olfactory receptors (ORs). We have selected the archaeon Haloferax volcanii as a cell host system and one of the most extensively investigated OR, namely I7-OR, whose pre...

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Detalles Bibliográficos
Autores principales: Lobasso, Simona, Vitale, Rita, Lopalco, Patrizia, Corcelli, Angela
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2015
Materias:
Communication
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4390878/
https://www.ncbi.nlm.nih.gov/pubmed/25761264
http://dx.doi.org/10.3390/life5010770
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author Lobasso, Simona
Vitale, Rita
Lopalco, Patrizia
Corcelli, Angela
author_facet Lobasso, Simona
Vitale, Rita
Lopalco, Patrizia
Corcelli, Angela
author_sort Lobasso, Simona
collection PubMed
description The aim of this study was to explore the possibility of using an archaeal microorganism as a host system for expressing mammalian olfactory receptors (ORs). We have selected the archaeon Haloferax volcanii as a cell host system and one of the most extensively investigated OR, namely I7-OR, whose preferred ligands are short-chain aldehydes, such as octanal, heptanal, nonanal. A novel plasmid has been constructed to express the rat I7-OR, fused with a hexahistidine-tag for protein immunodetection. The presence of the recombinant receptor at a membrane level was demonstrated by immunoblot of the membranes isolated from the transgenic archaeal strain. In addition, the lipid composition of archaeonanosomes containing ORs has been characterized in detail by High-Performance Thin-Layer Chromatography (HPTLC) in combination with Matrix-Assisted Laser Desorption Ionization—Time-Of-Flight/Mass Spectrometry (MALDI-TOF/MS) analysis.
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spelling pubmed-43908782015-05-21 Haloferax volcanii, as a Novel Tool for Producing Mammalian Olfactory Receptors Embedded in Archaeal Lipid Bilayer Lobasso, Simona Vitale, Rita Lopalco, Patrizia Corcelli, Angela Life (Basel) Communication The aim of this study was to explore the possibility of using an archaeal microorganism as a host system for expressing mammalian olfactory receptors (ORs). We have selected the archaeon Haloferax volcanii as a cell host system and one of the most extensively investigated OR, namely I7-OR, whose preferred ligands are short-chain aldehydes, such as octanal, heptanal, nonanal. A novel plasmid has been constructed to express the rat I7-OR, fused with a hexahistidine-tag for protein immunodetection. The presence of the recombinant receptor at a membrane level was demonstrated by immunoblot of the membranes isolated from the transgenic archaeal strain. In addition, the lipid composition of archaeonanosomes containing ORs has been characterized in detail by High-Performance Thin-Layer Chromatography (HPTLC) in combination with Matrix-Assisted Laser Desorption Ionization—Time-Of-Flight/Mass Spectrometry (MALDI-TOF/MS) analysis. MDPI 2015-03-09 /pmc/articles/PMC4390878/ /pubmed/25761264 http://dx.doi.org/10.3390/life5010770 Text en © 2015 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Communication
Lobasso, Simona
Vitale, Rita
Lopalco, Patrizia
Corcelli, Angela
Haloferax volcanii, as a Novel Tool for Producing Mammalian Olfactory Receptors Embedded in Archaeal Lipid Bilayer
title Haloferax volcanii, as a Novel Tool for Producing Mammalian Olfactory Receptors Embedded in Archaeal Lipid Bilayer
title_full Haloferax volcanii, as a Novel Tool for Producing Mammalian Olfactory Receptors Embedded in Archaeal Lipid Bilayer
title_fullStr Haloferax volcanii, as a Novel Tool for Producing Mammalian Olfactory Receptors Embedded in Archaeal Lipid Bilayer
title_full_unstemmed Haloferax volcanii, as a Novel Tool for Producing Mammalian Olfactory Receptors Embedded in Archaeal Lipid Bilayer
title_short Haloferax volcanii, as a Novel Tool for Producing Mammalian Olfactory Receptors Embedded in Archaeal Lipid Bilayer
title_sort haloferax volcanii, as a novel tool for producing mammalian olfactory receptors embedded in archaeal lipid bilayer
topic Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4390878/
https://www.ncbi.nlm.nih.gov/pubmed/25761264
http://dx.doi.org/10.3390/life5010770
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