Fluorescence techniques for fundamental and applied studies of membrane protein receptors: the 5-HT3 serotonin receptor
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Novel fluorescence approaches to investigate ligand recognition and structure of G protein-coupled receptors in native membranes have been developed. These methods combine the biosynthetic incorporation of unnatural fluorescent amino acids at known sites i ...
The screening of ligands for membrane receptor proteins is central to the discovery of new pharmaceutical drugs. We present a general method to reversibly attach receptor proteins via an affinity tag to a quartz surface and subsequently detect with high se ...
We have expressed the 5-HT3 receptor with two transient expression sytems: the Semliki Forest virus and the HEK293EBNA. Both systems allow the rapid expression of functional receptor protein at very high level: up to 107 receptors per cell. Large scale pro ...
The pore-forming toxin aerolysin is secreted by Aeromonas hydrophila as an inactive precursor. Based on chemical cross-linking and gel filtration, we show here that proaerolysin exists as a monomer at low concentrations but is dimeric above 0.1 mg/ml. At i ...
G protein-coupled receptors (GPCRs) constitute an abundant family of membrane receptors of high pharmacol. interest. Cell-based assays are the predominant means of assessing GPCR activation, but are limited by their inherent complexity. Functional mol. ass ...
A review with 6 refs. of the authors' work in developing a biophys. technique to investigate the structure, function and dynamics of membrane receptors. A fluorescence-based approach has been developed and applied to the prototypic G protein-coupled recept ...
Reversible and oriented immobilization of proteins in a functionally active form on solid surfaces is a prerequisite for the investigation of mol. interactions by surface-sensitive techniques. We demonstrate a method generally applicable for the attachment ...
The thermodn. and kinetics of ligand binding to the purified serotonin 5HT3 receptor and the local environment of the bound ligand were studied by fluorescence spectroscopy using a novel fluorescein-labeled ligand GR-flu [1,2,3,9-tetrahydro-3-[(5-methyl-1H ...
A fluorescent unnatural amino acid was introduced biosynthetically at known sites into the G protein-coupled neurokinin (tachykinin) NK2 receptor by suppression of UAG nonsense codons with the aid of a chem. misacylated synthetic tRNA specifically designed ...
A general method for understanding the mechanisms of ligand recognition and activation of G protein-coupled receptors has been developed. A study of ligand-receptor interactions in the prototypic seven-transmembrane neurokinin-2 receptor (NK2) using this f ...