ArticleProceedings of the National Academy of Sciences of the United States of America2009
Large-scale production and study of a synthetic G protein-coupled receptor: human olfactory receptor 17-4.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2009. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 46 papers.
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Who cites it
46 citing papers in PubMed, 101 citations in OpenAlex.
- Membrane Protein Folding and Biogenesis: Insights from Single-Molecule Force Spectroscopy.Chemical reviews · 2026Review
- Antagonists Enhance Cell-Surface Expression of Mammalian Odorant Receptors.International journal of molecular sciences · 2025Article
- Genome-wide pan-GPCR cell libraries accelerate drug discovery.Acta pharmaceutica Sinica. B · 2024Review
- Article
- Deciphering olfactory receptor binding mechanisms: a structural and dynamic perspective on olfactory receptors.Frontiers in molecular biosciences · 2024Review
- Article
- OR2AT4 and OR1A2 counterregulate molecular pathophysiological processes of steroid-resistant inflammatory lung diseases in human alveolar macrophages.Molecular medicine (Cambridge, Mass.) · 2022Article
- The Third Extracellular Loop of Mammalian Odorant Receptors Is Involved in Ligand Binding.International journal of molecular sciences · 2022Article
- OlfactionBase: a repository to explore odors, odorants, olfactory receptors and odorant-receptor interactions.Nucleic acids research · 2022Article
- Biophysical and functional characterization of the human TAS1R2 sweet taste receptor overexpressed in a HEK293S inducible cell line.Scientific reports · 2021Article
- An Overview of Artificial Olfaction Systems with a Focus on Surface Plasmon Resonance for the Analysis of Volatile Organic Compounds.Biosensors · 2021Review
- Thermostability of a recombinant G protein-coupled receptor expressed at high level in mammalian cell culture.Scientific reports · 2020Article
- Article
- Copper-mediated thiol potentiation and mutagenesis-guided modeling suggest a highly conserved copper-binding motif in human OR2M3.Cellular and molecular life sciences : CMLS · 2020Article
- The N-terminal region of RTP1S plays important roles in dimer formation and odorant receptor-trafficking.The Journal of biological chemistry · 2019Article
- G-Protein Coupled Receptor Protein Synthesis on a Lipid Bilayer Using a Reconstituted Cell-Free Protein Synthesis System.Life (Basel, Switzerland) · 2018Article
- Structure-Function Relationships of Olfactory and Taste Receptors.Chemical senses · 2018Review
- Quantification of interacting cognate odorants with olfactory receptors in nanovesicles.Scientific reports · 2017Article
- High-efficient production and biophysical characterisation of nicastrin and its interaction with APPC100.Scientific reports · 2017Article
- Medium-chain fatty acids modulate myocardial function via a cardiac odorant receptor.Basic research in cardiology · 2017Article
Corrections and comments
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Authors and funding
10 authors at 2 institutions in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
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 approximately 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.
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