ArticlePloS one2011
A robust and rapid method of producing soluble, stable, and functional G-protein coupled receptors.
Article in PloS one, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 19 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
19 citing papers in PubMed.
- Design and validation of recombinant protein standards for quantitative Western blot analysis of cannabinoid CBMicrobial cell factories · 2022Article
- Autoimmunity to Sphingosine-1-Phosphate-Receptors in Systemic Sclerosis and Pulmonary Arterial Hypertension.Frontiers in immunology · 2022Article
- Tips and tricks for cannabinoid receptor 1 detection, interaction and interpretation.Neural regeneration research · 2021Article
- The G protein coupled receptor CXCR4 designed by the QTY code becomes more hydrophilic and retains cell signaling activity.Scientific reports · 2020Article
- Antigen Design for Successful Isolation of Highly Challenging Therapeutic Anti-GPCR Antibodies.International journal of molecular sciences · 2020Review
- Self-assembling peptides: From a discovery in a yeast protein to diverse uses and beyond.Protein science : a publication of the Protein Society · 2020Article
- Cell-Free Expression and Photo-Crosslinking of the Human Neuropeptide YFrontiers in pharmacology · 2019Article
- G-Protein Coupled Receptor Protein Synthesis on a Lipid Bilayer Using a Reconstituted Cell-Free Protein Synthesis System.Life (Basel, Switzerland) · 2018Article
- Nanodelivery of a functional membrane receptor to manipulate cellular phenotype.Scientific reports · 2018Article
- Functional G-Protein-Coupled Receptor (GPCR) Synthesis: The Pharmacological Analysis of Human Histamine H1 Receptor (HRH1) Synthesized by a Wheat Germ Cell-Free Protein Synthesis System Combined with Asolectin Glycerosomes.Frontiers in pharmacology · 2018Article
- Production of G protein-coupled receptors in an insect-based cell-free system.Biotechnology and bioengineering · 2017Article
- Cell-free expression, purification, and ligand-binding analysis of Drosophila melanogaster olfactory receptors DmOR67a, DmOR85b and DmORCO.Scientific reports · 2015Article
- Stabilization of G protein-coupled receptors by point mutations.Frontiers in pharmacology · 2015Review
- Large-scale production and protein engineering of G protein-coupled receptors for structural studies.Frontiers in pharmacology · 2015Review
- Expression, surface immobilization, and characterization of functional recombinant cannabinoid receptor CB2.Biochimica et biophysica acta · 2013Article
- Lack of specificity shown by P2Y6 receptor antibodies.Naunyn-Schmiedeberg's archives of pharmacology · 2013Article
- Article
- Milligram production and biological activity characterization of the human chemokine receptor CCR3.PloS one · 2013Article
- Designer lipid-like peptides: a class of detergents for studying functional olfactory receptors using commercial cell-free systems.PloS one · 2011Article
Corrections and comments
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Authors and funding
12 authors.
Funding
Abstract
Membrane proteins, particularly G-protein coupled receptors (GPCRs), are notoriously difficult to express. Using commercial E. coli cell-free systems with the detergent Brij-35, we could rapidly produce milligram quantities of 13 unique GPCRs. Immunoaffinity purification yielded receptors at >90% purity. Secondary structure analysis using circular dichroism indicated that the purified receptors were properly folded. Microscale thermophoresis, a novel label-free and surface-free detection technique that uses thermal gradients, showed that these receptors bound their ligands. The secondary structure and ligand-binding results from cell-free produced proteins were comparable to those expressed and purified from HEK293 cells. Our study demonstrates that cell-free protein production using commercially available kits and optimal detergents is a robust technology that can be used to produce sufficient GPCRs for biochemical, structural, and functional analyses. This robust and simple method may further stimulate others to study the structure and function of 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.