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Heuristic Statistical Analysis of Fluorescence Fluctuation Data with Bright Spikes: Application to Ligand Binding to the Human Serotonin Receptor Expressed in Escherichia coli Cells
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Elke Van Craenenbroeck
Published/Copyright:
June 1, 2005
Abstract
A statistical method for the analysis of fluorescence fluctuation amplitudes including bright spikes is presented. This situation arises e. g. when fluorescent ligands interact with receptors carrying multiple binding sites. The technique gives information on the amount of bound ligand in solution, making it a complementary technique to fluorescence correlation spectroscopy analysis, which cannot be applied in this situation. Two simple statistical tests are proposed that can discriminate between fluorescence intensities originating from free ligands or complexes. The performance of the two tests is evaluated and compared on mixtures of a fluorophore and fluorophore coated beads that mimic the behaviour of multiliganded complexes. An application to ligand binding to the serotonin receptor, expressed on Escherichia coli cells, is also provided. Specific binding of a fluorophore to this receptor, as well as competition with several ligands, is assessed.
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Published Online: 2005-06-01
Published in Print: 2001-03-21
Copyright © 2001 by Walter de Gruyter GmbH & Co. KG
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Articles in the same Issue
- Highlight: Fluorescence Correlation Spectroscopy
- Heuristic Statistical Analysis of Fluorescence Fluctuation Data with Bright Spikes: Application to Ligand Binding to the Human Serotonin Receptor Expressed in Escherichia coli Cells
- Fluorescence Correlation Spectroscopy as a Tool to Investigate Single Molecule Probe Dynamics in Thin Polymer Films
- Ligand-Receptor Interactions in the Membrane of Cultured Cells Monitored by Fluorescence Correlation Spectroscopy
- Fluorescence Fluctuation Analysis for the Study of Interactions between Oligonucleotides and Polycationic Polymers
- Determination of the Net Exchange Rate of Tubulin Dimer in Steady-State Microtubules by Fluorescence Correlation Spectroscopy
- UV-Fluorescence Correlation Spectroscopy of 2-Aminopurine
- Isolation of Two cDNAs Encoding Functional Human Cytoplasmic Cysteinyl-tRNA Synthetase
- Involvement of Intracellular Ca2+in the Regulation of Bovine Leukemia Virus Expression
- Nucleotide-Binding Sites in the Functional Unit of Sarcoplasmic Reticulum Ca2+-ATPase as Studied by Photoaffinity Spin-Labeled 2-N3-SL-ATP
- Interaction between Lipopolysaccharide (LPS), LPS-Binding Protein (LBP), and Planar Membranes
- Differential Binding of Urokinase and Peptide Antagonists to the Urokinase Receptor: Evidence from Characterization of the Receptor in Four Primate Species
- Effects of Cysteine to Serine Substitutions in the Two Inter-Chain Disulfide Bonds of Insulin
- Tricorn Protease in Bacteria: Characterization of the Enzyme from Streptomyces coelicolor
- Structures of Tryparedoxins Revealing Interaction with Trypanothione
- Quantitative Two-Color Fluorescence Cross-Correlation Spectroscopy in the Analysis of Polymerase Chain Reaction
- Fluorescence Correlation Spectroscopy as a New Method for the Investigation of Aptamer/Target Interactions
- Localization of Lysobisphosphatidic Acid-Rich Membrane Domains in Late Endosomes
- Fluorescence Correlation Spectroscopy for the Characterisation of Drug Delivery Systems
- Accessing Molecular Dynamics in Cells by Fluorescence Correlation Spectroscopy
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