Publication

Large-scale production and study of a synthetic G protein-coupled receptor: Human olfactory receptor 17-4

Horst Vogel, Horst Pick
2009
Article
Résumé

Although understanding of the olfactory system has progressed at the level of downstream receptor signaling and the wiring of olfactory neurons, the system remains poorly understood at the molecular level of the receptors and their interaction with and recognition of odorant ligands. The structure and functional mechanisms of these receptors still remain a tantalizing enigma, because numerous previous attempts at the large-scale production of functional olfactory receptors (ORs) have not been successful to date. To investigate the elusive biochemistry and molecular mechanisms of olfaction, we have developed a mammalian expression system for the large-scale production and purification of a functional OR protein in milligram quantities. Here, we report the study of human OR17-4 (hOR17-4) purified from a HEK293S tetracycline-inducible system. Scale-up of production yield was achieved through suspension culture in a bioreactor, which enabled the preparation of > 10 mg of monomeric hOR17-4 receptor after immunoaffinity and size exclusion chromatography, with expression yields reaching 3 mg/L of culture medium. Several key post-translational modifications were identified using MS, and CD spectroscopy showed the receptor to be approximate to 50% alpha-helix, similar to other recently determined G protein-coupled receptor structures. Detergent-solubilized hOR17-4 specifically bound its known activating odorants lilial and floralozone in vitro, as measured by surface plasmon resonance. The hOR17-4 also recognized specific odorants in heterologous cells as determined by calcium ion mobilization. Our system is feasible for the production of large quantities of OR necessary for structural and functional analyses and research into OR biosensor devices.

À propos de ce résultat
Cette page est générée automatiquement et peut contenir des informations qui ne sont pas correctes, complètes, à jour ou pertinentes par rapport à votre recherche. Il en va de même pour toutes les autres pages de ce site. Veillez à vérifier les informations auprès des sources officielles de l'EPFL.
Proximité ontologique
Concepts associés (32)
Récepteur olfactif
vignette|redresse=1.5|Mécanismes de transduction des signaux odorants au niveau de la membrane des neurones récepteurs olfactifs. La liaison d'une molécule odorante à un récepteur olfactif (vert) modifie sa structure et active une protéine G (G). Celle-ci active à son tour l'adénylate cyclase (AC) augmentant la concentration d'AMPc intracellulaire qui, en se liant à des canaux ioniques, induit une dépolarisation du neurone récepteur olfactif.
Bulbe olfactif
Le bulbe olfactif (BO), parfois appelé lobe olfactif, est une région du cerveau des vertébrés dont la fonction principale est de traiter les informations olfactives en provenance des neurones chémorécepteurs olfactifs. C'est une structure paire – il y a deux bulbes olfactifs – légèrement détachée du reste du cerveau et la plus proche de la cavité nasale. Le bulbe olfactif est la première région du système nerveux central à traiter l'information olfactive.
Olfactory system
The olfactory system, or sense of smell, is the sensory system used for smelling (olfaction). Olfaction is one of the special senses, that have directly associated specific organs. Most mammals and reptiles have a main olfactory system and an accessory olfactory system. The main olfactory system detects airborne substances, while the accessory system senses fluid-phase stimuli. The senses of smell and taste (gustatory system) are often referred to together as the chemosensory system, because they both give the brain information about the chemical composition of objects through a process called transduction.
Afficher plus
Publications associées (40)

Olfactory modulation of barrel cortex activity during active whisking and passive whisker stimulation

Anthony Pierre Robert Renard

Rodents depend on olfaction and touch to meet many of their fundamental needs. However, the impact of simultaneous olfactory and tactile inputs on sensory representations in the cortex remains elusive. To study these interactions, we recorded large populat ...
NATURE PORTFOLIO2022

Microfluidic device performing on flow study of serial cell–cell interactions of two cell populations

Philippe Renaud, George Coukos, Thamani Dahoun, Margaux Catherine Marie Duchamp, Alexandre Harari, Marion Arnaud

In this study we present a novel microfluidic hydrodynamic trapping device to probe the cell–cell interaction between all cell samples of two distinct populations. We have exploited an hydrodynamic trapping method using microfluidics to immobilize a batch ...
2019

Computational modeling of the olfactory receptor Olfr73 suggests a molecular basis for low potency of olfactory receptor-activating compounds

Horst Vogel, Horst Pick, Thamani Dahoun, Shuguang Yuan, Marc Brugarolas Campillos

The mammalian olfactory system uses hundreds of specialized G-protein-coupled olfactory receptors (ORs) to discriminate a nearly unlimited number of odorants. Cognate agonists of most ORs have not yet been identified and potential non-olfactory processes m ...
Springer2019
Afficher plus
MOOCs associés (9)
Introduction à l'immunologie (part 1)
Ce cours décrit les mécanismes fondamentaux du système immunitaire pour mieux comprendre les bases immunologiques dela vaccination, de la transplantation, de l’immunothérapie, de l'allergie et des mal
Afficher plus

Graph Chatbot

Chattez avec Graph Search

Posez n’importe quelle question sur les cours, conférences, exercices, recherches, actualités, etc. de l’EPFL ou essayez les exemples de questions ci-dessous.

AVERTISSEMENT : Le chatbot Graph n'est pas programmé pour fournir des réponses explicites ou catégoriques à vos questions. Il transforme plutôt vos questions en demandes API qui sont distribuées aux différents services informatiques officiellement administrés par l'EPFL. Son but est uniquement de collecter et de recommander des références pertinentes à des contenus que vous pouvez explorer pour vous aider à répondre à vos questions.