beta 1-adrenergic receptor association with PSD-95. Inhibition of receptor internalization and facilitation of beta 1-adrenergic receptor interaction with N-methyl-D-aspartate receptors.
The beta(1)-adrenergic receptor (beta(1)AR) is the most abundant subtype of beta-adrenergic receptor in the mammalian brain and is known to potently regulate synaptic plasticity. To search for potential neuronal beta(1)AR-interacting proteins, we screened a rat brain cDNA library using the beta(1)AR carboxyl terminus (beta(1)AR-CT) as bait in the yeast two-hybrid ... system. These screens identified PSD-95, a multiple PDZ domain-containing scaffolding protein, as a specific binding partner of the beta(1)AR-CT. This interaction was confirmed by in vitro fusion protein pull-down and blot overlay experiments, which demonstrated that the beta(1)AR-CT binds specifically to the third PDZ domain of PSD-95. Furthermore, the full-length beta(1)AR associates with PSD-95 in cells, as determined by co-immunoprecipitation experiments and immunofluorescence co-localization studies. The interaction between beta(1)AR and PSD-95 is mediated by the last few amino acids of the beta(1)AR, and mutation of the beta(1)AR carboxyl terminus eliminated the binding and disrupted the co-localization of the beta(1)AR and PSD-95 in cells. Agonist-induced internalization of the beta(1)AR in HEK-293 cells was markedly attenuated by PSD-95 co-expression, whereas co-expression of PSD-95 has no significant effect on either desensitization of the beta(1)AR or beta(1)AR-induced cAMP accumulation. Furthermore, PSD-95 facilitated the formation of a complex between the beta(1)AR and N-methyl-d-aspartate receptors, as assessed by co-immunoprecipitation. These data reveal that PSD-95 is a specific beta(1)AR binding partner that modulates beta(1)AR function and facilitates physical association of the beta(1)AR with synaptic proteins, such as the N-methyl-d-aspartate receptors, which are known to be regulated by beta(1)AR stimulation.
Mesh Terms:
Animals, Brain, COS Cells, Cell Line, Cercopithecus aethiops, Cloning, Molecular, Gene Library, Humans, Intracellular Signaling Peptides and Proteins, Membrane Proteins, Nerve Tissue Proteins, Peptide Fragments, Protein Binding, Protein Transport, Rats, Receptors, Adrenergic, beta-1, Receptors, N-Methyl-D-Aspartate, Recombinant Fusion Proteins, Saccharomyces cerevisiae, Transfection
Animals, Brain, COS Cells, Cell Line, Cercopithecus aethiops, Cloning, Molecular, Gene Library, Humans, Intracellular Signaling Peptides and Proteins, Membrane Proteins, Nerve Tissue Proteins, Peptide Fragments, Protein Binding, Protein Transport, Rats, Receptors, Adrenergic, beta-1, Receptors, N-Methyl-D-Aspartate, Recombinant Fusion Proteins, Saccharomyces cerevisiae, Transfection
J. Biol. Chem.
Date: Dec. 08, 2000
PubMed ID: 10995758
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