Dual activities of odorants on olfactory and nuclear hormone receptors.

Pick, Horst; Etter, Sylvain; Baud, Olivia; Schmauder, Ralf; Bordoli, Lorenza; Schwede, Torsten; Vogel, Horst

doi:10.1074/jbc.a109.040964

Cited by 3 publications

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“…Studies focusing on other GPCRs, like rhodopsin and the AT1 angiotensin receptor, have shown that loss of glycosylation may lead to a decreased receptor function or stability. , These findings indicate that the production of ORs in mammalian cells might be crucial for functional expression and purification. Apart from these obvious advantages, mammalian expression systems also offer the possibility of probing OR function directly in the same heterologous cell type used for protein production. ,,, This is not possible in bacterial or insect cell expression systems or in the recently emerging cell-free OR production systems. − …”

Section: Discussionmentioning

confidence: 99%

“…Furthermore, no high-resolution three-dimensional (3D) structure of an OR has been determined, in contrast to those of other mammalian GPCRs. − Consequently, a reliable structural basis of OR’s ligand recognition is unknown. Therefore, only models of ligand binding sites of ORs have been proposed on the basis of sequence comparisons between ORs, by functional studies of mutant OR proteins, and by comparisons with known 3D structures of other GPCRs. − Finally, quantitative thermodynamic and kinetic constants for interactions between ORs and their activating ligands or their downstream signaling proteins are totally missing. Hence, methods for the efficient production of ORs in their biologically active form are of crucial importance.…”

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confidence: 99%

“…As no universal expression system has been discovered yet for the functional production of ORs, the proper combination of target receptor and expression host must be found empirically for each case . HEK293 cells have been shown to be suitable for the functional expression of a few ORs ,,, and also for large-scale OR production in one case …”

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confidence: 99%

“…31 As no universal expression system has been discovered yet for the functional production of ORs, the proper combination of target receptor and expression host must be found empirically for each case. 32 functional expression of a few ORs 19,20,33,34 and also for large-scale OR production in one case. 35 Here, we achieved by transient transfection a considerably high yield of functional OR cell surface expression of mouse mOR256-17 in HEK293 cells.…”

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Recombinant Expression and Functional Characterization of Mouse Olfactory Receptor mOR256-17 in Mammalian Cells

et al. 2011

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Olfactory receptors (ORs) constitute the largest family of sensory membrane proteins in mammals. They play a key role within the olfactory system in recognizing and discriminating a nearly unlimited number of structurally diverse odorous molecules. The molecular basis of OR-mediated signal detection and transduction is poorly understood. This is due to difficulties in functional expression of ORs in high yields, preventing structural and biophysical studies at the level of the receptor protein. Here we report on recombinant expression of mouse receptor mOR256-17 yielding 10(6) ORs per cell in transiently transfected mammalian cells. For quantification and optimization of OR expression, we employed different fluorescent probes. Green fluorescent protein fused to the C-terminus of mOR256-17 allowed quantification of total cellular OR biosynthesis, and post-translational fluorescence labeling of a 12-amino acid polypeptide sequence at the N-terminus permitted the selective visualization and quantification of ORs at the plasma membrane using cell flow cytometry. Our dual-color labeling approach is generally applicable to quantification of membrane proteins for mammalian cell-based expression. By screening a large odorant compound library, we discovered a selective spectrum of potent mOR256-17-specific agonists essential for probing the receptor function for future scaled-up productions.

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Section: Discussionmentioning

confidence: 99%

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Recombinant Expression and Functional Characterization of Mouse Olfactory Receptor mOR256-17 in Mammalian Cells

et al. 2011

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“…The expression of some ORs is reported to be controlled by odorants which are dually active; OR2A1 is known to be regulated by sandalwood compounds which initiate neuronal signaling as well as hormonal transcriptional control of speci c genes. [45,44]. OR51E2/PSGR, a marker of prostate cancer and OR7D4 can be activated by androstenone [46][47][48][49][50], also, OR51E1 is reported to associate with gastrointestinal neuroendocrine carcinomas [51], It can be activated by steroid hormones [13].…”

Section: Discussionmentioning

confidence: 99%

Breast Cancer and Smell: Hints from Epigenetic and Functional Alteration of the Olfaction

Fessahaye

Khalid

Mohamed

et al. 2021

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Purpose: Olfactory receptors are G protein coupled surface receptors (GPCRs) of which their ectopic expression is currently of mounting interest to the development and metastasis of malignancies. These genes having a direct contact with the environment may probably be stimulated by various factors which can bring about methylation aberrations, including DNA hypo and hyper-methylation. Here we gather clues from epigenetic and phenotypic data in order to further our understanding of the potential association of the olfaction with oncogenesis.Methods: Whole methylome dataset of breast cancer series generated by Illumina Infinium Human Methylation 450 Bead Chip was interrogated for differentially methylated genes and further subject to network analysis using various search tools. Analysis of putative phenotypic trait in olfaction function was performed using smell detection and smell identification tests and data was analyzed using Mann-Whitney test.Results: Sixty-eight differentially methylated ORs were enriched mainly on chromosomes 1q23, and 11p15, specifically 1q44 (P value 6.867e-20). Amongst the disease signatures of these hypomethylation events was breast cancer itself (P value 0.004437). Network analysis suggests the interaction of differentially hypo and hyper methylated olfactory receptor genes might be pivotal in stimulating several important biological pathways including circadian genes and pathways potentially associated with metastasis. Phenotypic smell test shows a generalized impairment of smell capability in breast cancer patients as compared to controls (Mann-Whitney Test P=0.0001), an effect that is independent of chemotherapyConclusions: The olfaction appears as a crucial element in carcinogenesis, evident by both phenotypic and genotypic (epigenetic) data in a well characterized breast cancer subset.

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The Mouse Eugenol Odorant Receptor: Structural and Functional Plasticity of a Broadly Tuned Odorant Binding Pocket

et al. 2010

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Molecular interactions of odorants with their olfactory receptors (ORs) are of central importance for the ability of the mammalian olfactory system to detect and discriminate a vast variety of odors with a limited set of receptors. How a particular OR binds and distinguishes different odorant molecules remains largely unknown on a structural basis. Here we investigated this question for the mouse eugenol receptor (mOR-EG). By screening a large odorant library, we discovered a wide range of chemical structures activating the receptor in heterologous mammalian cells. Potent agonists comprise (i) benzene, (ii) cyclohexane, or (iii) polycyclic structures substituted with alcohol, aldehyde, keto, ether, or esterified carboxylic groups. To detect those amino acids within the receptor that are in contact with a particular bound odorant molecule, we investigated how distinct mOR-EG point mutants were activated by the different odorant agonists found for the wild-type receptor. We identified 11 amino acids as a part of the receptor's ligand binding pocket. Molecular modeling predicted 10 of these residues in transmembrane helices TM3-TM6 and one in the extracellular loop between TM2 and TM3. These amino acids participate in odorant binding with variable importance depending on the type of odorant, revealing functional "fingerprints" of ligand-receptor interactions.

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Dual activities of odorants on olfactory and nuclear hormone receptors.

Abstract: On page 30551, Table 1, first column, the experimentally determined values of EC 50 of ER activation for MC, Polysantol, Javanol, and androstenol have been presented with the wrong concentration (mM). The correct concentration is M.

Cited by 3 publications

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Recombinant Expression and Functional Characterization of Mouse Olfactory Receptor mOR256-17 in Mammalian Cells

Recombinant Expression and Functional Characterization of Mouse Olfactory Receptor mOR256-17 in Mammalian Cells

Breast Cancer and Smell: Hints from Epigenetic and Functional Alteration of the Olfaction

The Mouse Eugenol Odorant Receptor: Structural and Functional Plasticity of a Broadly Tuned Odorant Binding Pocket

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