ArticleProceedings of the National Academy of Sciences of the United States of America2008
Efficient cell-free production of olfactory receptors: detergent optimization, structure, and ligand binding analyses.
Article in Proceedings of the National Academy of Sciences of the United States of America, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 44 papers.
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Who cites it
44 citing papers in PubMed, 134 citations in OpenAlex.
- Portable geosmin detection system based on sensor cells expressing insect odorant receptors.Scientific reports · 2026Article
- Half a Century of Progress: The Evolution of Wheat Germ-Based In Vitro Translation into a Versatile Protein Production Method.International journal of molecular sciences · 2025Review
- Cell-free protein synthesis platforms for accelerating drug discovery.Biotechnology notes (Amsterdam, Netherlands) · 2025Review
- Biosensors for Odor Detection: A Review.Biosensors · 2023Review
- Enabling technology and core theory of synthetic biology.Science China. Life sciences · 2023Review
- Transcuticular calcium imaging as a tool for the functional study of insect odorant receptors.Frontiers in molecular neuroscience · 2023Article
- Field-Effect Sensors Using Biomaterials for Chemical Sensing.Sensors (Basel, Switzerland) · 2021Review
- Easy Synthesis of Complex Biomolecular Assemblies: Wheat Germ Cell-Free Protein Expression in Structural Biology.Frontiers in molecular biosciences · 2021Review
- Bio-Inspired Strategies for Improving the Selectivity and Sensitivity of Artificial Noses: A Review.Sensors (Basel, Switzerland) · 2020Review
- Structural instability and divergence from conserved residues underlie intracellular retention of mammalian odorant receptors.Proceedings of the National Academy of Sciences of the United States of America · 2020Article
- Review
- Synthetic Biology Goes Cell-Free.BMC biology · 2019Review
- Human-like smelling of a rose scent using an olfactory receptor nanodisc-based bioelectronic nose.Scientific reports · 2018Article
- Single-pot glycoprotein biosynthesis using a cell-free transcription-translation system enriched with glycosylation machinery.Nature communications · 2018Article
- Applications and Advances in Bioelectronic Noses for Odour Sensing.Sensors (Basel, Switzerland) · 2018Review
- Cell-free production and characterisation of human uncoupling protein 1-3.Biochemistry and biophysics reports · 2017Article
- The role of metals in mammalian olfaction of low molecular weight organosulfur compounds.Natural product reports · 2017Review
- Preservation of protein expression systems at elevated temperatures for portable therapeutic production.Journal of the Royal Society, Interface · 2017Article
- Cell-Free Synthetic Biology: Engineering Beyond the Cell.Cold Spring Harbor perspectives in biology · 2016Review
- Cell-free translation and purification of Arabidopsis thaliana regulator of G signaling 1 protein.Protein expression and purification · 2016Article
Corrections and comments
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Authors and funding
6 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
High-level production of membrane proteins, particularly of G protein-coupled receptors (GPCRs) in heterologous cell systems encounters a number of difficulties from their inherent hydrophobicity in their transmembrane domains, which frequently cause protein aggregation and cytotoxicity and thus reduce the protein yield. Recent advances in cell-free protein synthesis circumvent those problems to produce membrane proteins with a yield sometimes exceeding the cell-based approach. Here, we report cell-free production of a human olfactory receptor 17-4 (hOR17-4) using the wheat germ extract. Using the simple method, we also successful produced two additional olfactory receptors. To obtain soluble olfactory receptors and to increase yield, we directly added different detergents in varying concentrations to the cell-free reaction. To identify a purification buffer system that maintained the receptor in a nonaggregated form, we developed a method that uses small-volume size-exclusion column chromatography combined with rapid and sensitive dot-blot detection. Different buffer components including salt concentration, various detergents and detergent concentration, and reducing agent and its concentrations were evaluated for their ability to maintain the cell-free produced protein stable and nonaggregated. The purified olfactory receptor displays a typical a alpha-helical CD spectrum. Surface plasmon resonance measurements were used to show binding of a known ligand undecanal to hOR17-4. Our approach to produce a high yield of purified olfactory receptor is a milestone toward obtaining a large quantity of olfactory receptors for designing bionic sensors. Furthermore, this simple approach may be broadly useful not only for other classes of GPCRs but also for other membrane proteins.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.