ArticleBiophysical journal2003
Confocal FRET microscopy to measure clustering of ligand-receptor complexes in endocytic membranes.
Article in Biophysical journal, 2003. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 43 papers.
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Who cites it
43 citing papers in PubMed, 102 citations in OpenAlex.
- Protein-protein interaction detection in plants: from classical approaches to future perspectives.Biologia futura · 2026Review
- In vivo quantitative FRET small animal imaging: Intensity versus lifetime-based FRET.Biophysical reports · 2023Article
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- MOP and NOP receptor interaction: Studies with a dual expression system and bivalent peptide ligands.PloS one · 2022Article
- Macroscopic Fluorescence Lifetime Imaging for Monitoring of Drug-Target Engagement.Methods in molecular biology (Clifton, N.J.) · 2022Article
- Quantitative FRET (qFRET) Technology for the Determination of Protein-Protein Interaction Affinity in Solution.Molecules (Basel, Switzerland) · 2021Review
- Milestones in the development and implementation of FRET-based sensors of intracellular signals: A biological perspective of the history of FRET.Cellular signalling · 2020Review
- Multiplexed non-invasive tumor imaging of glucose metabolism and receptor-ligand engagement using dark quencher FRET acceptor.Theranostics · 2020Article
- Association of microtubules and axonal RNA transferred from myelinating Schwann cells in rat sciatic nerve.PloS one · 2020Article
- Nociceptin/Orphanin FQ (N/OFQ) conjugated to ATTO594: a novel fluorescent probe for the N/OFQ (NOP) receptor.British journal of pharmacology · 2018Article
- TOWARD BAYESIAN INFERENCE OF THE SPATIAL DISTRIBUTION OF PROTEINS FROM THREE-CUBE FÖRSTER RESONANCE ENERGY TRANSFER DATA.The annals of applied statistics · 2017Article
- Pooled screening for antiproliferative inhibitors of protein-protein interactions.Nature chemical biology · 2016Article
- A new trend to determine biochemical parameters by quantitative FRET assays.Acta pharmacologica Sinica · 2015Review
- Antigen-B Cell Receptor Complexes Associate with Intracellular major histocompatibility complex (MHC) Class II Molecules.The Journal of biological chemistry · 2015Article
- Single-molecule analyses of fully functional fluorescent protein-tagged follitropin receptor reveal homodimerization and specific heterodimerization with lutropin receptor.Biology of reproduction · 2015Article
- Establishing a cellular FRET-based fluorescence plate reader assay to monitor proNGF-induced cross-linking of sortilin and the neurotrophin receptor p75(NTR).International journal of biochemistry and molecular biology · 2015Article
- Automated selection of regions of interest for intensity-based FRET analysis of transferrin endocytic trafficking in normal vs. cancer cells.Methods (San Diego, Calif.) · 2014Article
- Spatial light modulator based active wide-field illumination for ex vivo and in vivo quantitative NIR FRET imaging.Biomedical optics express · 2014Article
- IQGAP1 interactome analysis by in vitro reconstitution and live cell 3-color FRET microscopy.Cytoskeleton (Hoboken, N.J.) · 2013Article
- Intensity correlation-based calibration of FRET.Biophysical journal · 2013Article
Corrections and comments
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
The dynamics of protein distribution in endocytic membranes are relevant for many cellular processes, such as protein sorting, organelle and membrane microdomain biogenesis, protein-protein interactions, receptor function, and signal transduction. We have developed an assay based on Fluorescence Resonance Energy Microscopy (FRET) and novel mathematical models to differentiate between clustered and random distributions of fluorophore-bound molecules on the basis of the dependence of FRET intensity on donor and acceptor concentrations. The models are tailored to extended clusters, which may be tightly packed, and account for geometric exclusion effects between membrane-bound proteins. Two main criteria are used to show that labeled polymeric IgA-ligand-receptor complexes are organized in clusters within apical endocytic membranes of polarized MDCK cells: 1), energy transfer efficiency (E%) levels are independent of acceptor levels; and 2), with increasing unquenched donor: acceptor ratio, E% decreases. A quantitative analysis of cluster density indicates that a donor-labeled ligand-receptor complex should have 2.5-3 labeled complexes in its immediate neighborhood and that clustering may occur at a limited number of discrete membrane locations and/or require a specific protein that can be saturated. Here, we present a new sensitive FRET-based method to quantify the co-localization and distribution of ligand-receptor complexes in apical endocytic membranes of polarized cells.
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