Selective Interactions between Helix VIII of the Human μ-Opioid Receptors and the C Terminus of Periplakin Disrupt G Protein Activation
| Autor: | Graeme Milligan, Elaine Kellett, Giu-Jie Feng, Jonathan I. Wilde, Carol A. Scorer, Julia H. White |
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| Rok vydání: | 2003 |
| Předmět: |
Central Nervous System
DNA Complementary G protein Recombinant Fusion Proteins Immunoblotting Molecular Sequence Data Receptors Opioid mu Biotin Guanosine Plasma protein binding Biology Ligands Transfection Biochemistry Cell Line chemistry.chemical_compound Protein structure GTP-Binding Proteins Two-Hybrid System Techniques Humans Protein Isoforms Histidine Tissue Distribution Amino Acid Sequence RNA Messenger Receptor Molecular Biology Periplakin Gene Library Glutathione Transferase Microscopy Confocal Sequence Homology Amino Acid C-terminus Cell Membrane Plakins Brain DNA Cell Biology Fusion protein Molecular biology Actins Protein Structure Tertiary Cell biology Cytoskeletal Proteins Microscopy Fluorescence chemistry Guanosine 5'-O-(3-Thiotriphosphate) Protein Binding |
| Zdroj: | Journal of Biological Chemistry. 278:33400-33407 |
| ISSN: | 0021-9258 |
| DOI: | 10.1074/jbc.m305866200 |
| Popis: | Analysis of interactions between the C-terminal tail of the MOP-1 and MOP-1A variants of the human mu-opioid receptor with proteins derived from a human brain cDNA library resulted in identification of the actin and intermediate filament-binding protein periplakin. Mapping of this interaction indicated that the predicted fourth intracellular loop/helix VIII of the receptor interacts with the C-terminal rod and linker region of periplakin. Periplakin is widely expressed in the central nervous system of both man and rat and demonstrated an overlapping but not identical distribution with mu-opioid (MOP) receptors. Co-expression of periplakin with MOP-1 or a MOP-1-eYFP fusion construct in HEK293 cells did not interfere with agonist-mediated internalization of the receptor. When co-expressed with a MOP-1-Gi1 alpha fusion protein periplakin significantly reduced the capacity of the agonist to stimulate binding of 35S-labeled guanosine 5'-3-O-(thio)triphosphate ([35S]GTP gamma S) to the receptor-associated G protein. By contrast, periplakin did not interfere with agonist-stimulation of [35S]GTP gamma S binding to either an alpha 2A-adrenoreceptor-Gi1 alpha fusion protein or a beta2-adrenoreceptor-Gs alpha fusion protein, indicating its selectivity of function. This represents the first example of an opioid receptor-interacting protein that functions to disrupt agonist-mediated G protein activation. |
| Databáze: | OpenAIRE |
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