ArticleChembiochem : a European journal of chemical biology2014
Bioorthogonal fluorescent labeling of functional G-protein-coupled receptors.
Article in Chembiochem : a European journal of chemical biology, 2014. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 20 papers.
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Who cites it
20 citing papers in PubMed, 48 citations in OpenAlex.
- Noncanonical Amino Acid Tools and Their Application to Membrane Protein Studies.Chemical reviews · 2024Review
- Investigating Protein Degradability through Site-Specific Ubiquitin Ligase Recruitment.bioRxiv : the preprint server for biology · 2024Article
- Article
- Single-molecule FRET uncovers hidden conformations and dynamics of human Argonaute 2.Nature communications · 2022Article
- FRET sensors reveal the retinal entry pathway in the G protein-coupled receptor rhodopsin.iScience · 2022Article
- Frizzled BRET sensors based on bioorthogonal labeling of unnatural amino acids reveal WNT-induced dynamics of the cysteine-rich domain.Science advances · 2021Article
- Bioorthogonal Reactions of Triarylphosphines and Related Analogues.Chemical reviews · 2021Review
- A Genetically Encoded Picolyl Azide for Improved Live Cell Copper Click Labeling.Frontiers in chemistry · 2021Article
- Using genetically incorporated unnatural amino acids to control protein functions in mammalian cells.Essays in biochemistry · 2019Review
- Evaluation and optimisation of unnatural amino acid incorporation and bioorthogonal bioconjugation for site-specific fluorescent labelling of proteins expressed in mammalian cells.Biochemistry and biophysics reports · 2019Article
- Low- q Bicelles Are Mixed Micelles.The journal of physical chemistry letters · 2018Article
- Spirohexene-Tetrazine Ligation Enables Bioorthogonal Labeling of Class B G Protein-Coupled Receptors in Live Cells.Journal of the American Chemical Society · 2017Article
- The Energetics of Chromophore Binding in the Visual Photoreceptor Rhodopsin.Biophysical journal · 2017Article
- Measurement of Slow Spontaneous Release of 11-cis-Retinal from Rhodopsin.Biophysical journal · 2017Article
- Purification of family B G protein-coupled receptors using nanodiscs: Application to human glucagon-like peptide-1 receptor.PloS one · 2017Article
- A simple method for enhancing the bioorthogonality of cyclooctyne reagent.Chemical communications (Cambridge, England) · 2016Article
- Micelle-Enhanced Bioorthogonal Labeling of Genetically Encoded Azido Groups on the Lipid-Embedded Surface of a GPCR.Chembiochem : a European journal of chemical biology · 2015Article
- Conformational selection and equilibrium governs the ability of retinals to bind opsin.The Journal of biological chemistry · 2015Article
- Ghrelin receptor conformational dynamics regulate the transition from a preassembled to an active receptor:Gq complex.Proceedings of the National Academy of Sciences of the United States of America · 2015Article
- Multiplex detection of functional G protein-coupled receptors harboring site-specifically modified unnatural amino acids.Biochemistry · 2015Article
Corrections and comments
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Authors and funding
6 authors at 2 institutions in 2 countries.
Funding
Abstract
Novel methods are required for site-specific, quantitative fluorescent labeling of G-protein-coupled receptors (GPCRs) and other difficult-to-express membrane proteins. Ideally, fluorescent probes should perturb the native structure and function as little as possible. We evaluated bioorthogonal reactions to label genetically encoded p-acetyl-L-phenylalanine (AcF) or p-azido-L-phenylalanine (azF) residues in receptors heterologously expressed in mammalian cells. We found that keto-selective reagents were not truly bioorthogonal, possibly owing to post-translational protein oxidation reactions. In contrast, the strain-promoted [3+2] azide-alkyne cycloaddition (SpAAC) with dibenzocyclooctyne (DIBO) reagents yielded stoichiometric conjugates with azF-rhodopsin while undergoing negligible background reactions. As one application of this technique, we used Alexa488-rhodopsin to measure the kinetics of ligand uptake and release in membrane-mimetic bicelles using a novel fluorescence-quenching assay.
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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.