A disulfide bond between conserved cysteines in the extracellular loops of the human VIP receptor is required for binding and activation
| Autor: | Sanne Møller Knudsen, Jeppe W. Tams, Birgitte Schjellerup Wulff, Jan Fahrenkrug |
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| Rok vydání: | 1997 |
| Předmět: |
G protein
Biophysics Transfection Binding Competitive Biochemistry Protein Structure Secondary Dithiothreitol Cell Line Structure-Activity Relationship chemistry.chemical_compound Structural Biology Cyclic AMP Genetics Humans G protein-coupled receptor Cysteine Disulfides Receptor Site-directed mutagenesis Molecular Biology Vasoactive intestinal peptide receptor Mutagenesis Wild type Cell Biology chemistry Mutagenesis Site-Directed Receptors Vasoactive Intestinal Peptide Vasoactive Intestinal Peptide |
| Zdroj: | FEBS Letters. 412:141-143 |
| ISSN: | 0014-5793 |
| DOI: | 10.1016/s0014-5793(97)00714-x |
| Popis: | The importance of two highly conserved cysteines in the human vasoactive intestinal peptide receptor 1 (hVIPR 1) was examined. By site-directed mutagenesis each Cys residue was converted into Ala or Ser. The mutant and wild-type genes were transfected into HEK293 cells and tested for the ability to bind VIP and to activate cAMP production. Cys215-Ala/Ser and Cys285-Ala/Ser showed at least a 10-fold decrease in binding affinity and receptor potency when compared to the wild type. In contradiction to the wild-type receptor, both mutations were insensitive to dithiothreitol (DTT). The results indicate the existence of a disulfide bond between Cys215 and Cys285, which is important for stabilising the receptor in the correct conformation for ligand binding and activation. © 1997 Federation of European Biochemical Societies. |
| Databáze: | OpenAIRE |
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