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Identification by site-directed mutagenesis of amino acids contributing to ligand-binding specificity or signal transduction properties of the human FP prostanoid receptor
- Source :
- The Biochemical journal. 371(Pt 2)
- Publication Year :
- 2003
-
Abstract
- Prostanoid receptors belong to the class of heptahelical plasma membrane receptors. For the five prostanoids, eight receptor subtypes have been identified. They display an overall sequence similarity of roughly 30%. Based on sequence comparison, single amino acids in different subtypes of different species have previously been identified by site-directed mutagenesis or in hybrid receptors that appear to be essential for ligand binding or G-protein coupling. Based on this information, a series of mutants of the human FP receptor was generated and characterized in ligand-binding and second-messenger-formation studies. It was found that mutation of His-81 to Ala in transmembrane domain 2 and of Arg-291 to Leu in transmembrane domain 7, which are putative interaction partners for the prostanoid's carboxyl group, abolished ligand binding. Mutants in which Ser-263 in transmembrane domain 6 or Asp-300 in transmembrane domain 7 had been replaced by Ala or Gln, respectively, no longer discriminated between prostaglandins PGF2α and PGD2. Thus distortion of the topology of transmembrane domains 6 and 7 appears to interfere with the cyclopentane ring selectivity of the receptor. PGF2α-induced inositol formation was strongly reduced in the mutant Asp-300Gln, inferring a role for this residue in agonist-induced G-protein activation.
- Subjects :
- Placenta
Mutant
Receptors, Prostaglandin
Biology
Dinoprost
Ligands
Biochemistry
Cell surface receptor
Pregnancy
Humans
Site-directed mutagenesis
Receptor
Molecular Biology
DNA Primers
chemistry.chemical_classification
Binding Sites
Base Sequence
Mutagenesis
Cell Biology
Recombinant Proteins
Amino acid
Transmembrane domain
Kinetics
chemistry
Amino Acid Substitution
Mutagenesis, Site-Directed
Institut für Ernährungswissenschaft
Female
Signal transduction
Signal Transduction
Research Article
Subjects
Details
- ISSN :
- 02646021
- Volume :
- 371
- Issue :
- Pt 2
- Database :
- OpenAIRE
- Journal :
- The Biochemical journal
- Accession number :
- edsair.doi.dedup.....c0ff9dc3e603c2464bf79b0fbe0e2517