PDZ domain-mediated interactions of G protein-coupled receptors with postsynaptic density protein 95: quantitative characterization of interactions
| Autor: | Birgitte P.S. Jacky, Volker F. Wirth, Thor C. Møller, Karen L. Martinez, Julia Bender, Kristian Strømgaard, Nina I. Roberts, Anders Bach, Jan M. Deussing, Thue W. Schwartz |
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| Rok vydání: | 2013 |
| Předmět: |
Scaffold protein
Mouse PDZ Domains Hippocampus Biochemistry Receptors G-Protein-Coupled Substrate Specificity Chemokine receptor Mice Molecular Cell Biology Signaling in Cellular Processes Membrane Receptor Signaling Biomacromolecule-Ligand Interactions Neurons Multidisciplinary Intracellular Signaling Peptides and Proteins Animal Models Hormone Receptor Signaling Transport protein Cell biology Protein Transport Chemistry Medicine Disks Large Homolog 4 Protein Protein Binding Research Article Signal Transduction Science PDZ domain Biophysics Biology Protein–protein interaction Model Organisms Chemical Biology Animals Humans Amino Acid Sequence Protein Interactions G protein-coupled receptor Membrane Proteins Proteins Transmembrane Proteins Kinetics G-Protein Signaling HEK293 Cells Membrane protein Peptidomimetics Postsynaptic density |
| Zdroj: | PLoS ONE PLoS ONE, Vol 8, Iss 5, p e63352 (2013) Møller, T C, Wirth, V F, Roberts, N I, Bender, J, Bach, A, Jacky, B P S, Strømgaard, K, Deussing, J M, Schwartz, T W & Martinez, K L 2013, ' PDZ domain-mediated interactions of G protein-coupled receptors with postsynaptic density protein 95 : quantitative characterization of interactions ', PLOS ONE, vol. 8, no. 5, e63352 . https://doi.org/10.1371/journal.pone.0063352 PLoS ONE 8:e63352 (2013) |
| ISSN: | 1932-6203 |
| DOI: | 10.1371/journal.pone.0063352 |
| Popis: | G protein-coupled receptors (GPCRs) constitute the largest family of membrane proteins in the human genome. Their signaling is regulated by scaffold proteins containing PDZ domains, but although these interactions are important for GPCR function, they are still poorly understood. We here present a quantitative characterization of the kinetics and affinity of interactions between GPCRs and one of the best characterized PDZ scaffold proteins, postsynaptic density protein 95 (PSD-95), using fluorescence polarization (FP) and surface plasmon resonance (SPR). By comparing these in vitro findings with colocalization of the full-length proteins in cells and with previous studies, we suggest that the range of relevant interactions might extend to interactions with K-i = 450 mu M in the in vitro assays. Within this range, we identify novel PSD-95 interactions with the chemokine receptor CXCR2, the neuropeptide Y receptor Y-2, and four of the somatostatin receptors (SSTRs). The interaction with SSTR1 was further investigated in mouse hippocampal neurons, where we found a clear colocalization between the endogenously expressed proteins, indicating a potential for further investigation of the role of this interaction. The approach can easily be transferred to other receptors and scaffold proteins and this could help accelerate the discovery and quantitative characterization of GPCR-PDZ interactions. |
| Databáze: | OpenAIRE |
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