ArticleMolecular & cellular proteomics : MCP2008
A generic approach for the purification of signaling complexes that specifically interact with the carboxyl-terminal domain of G protein-coupled receptors.
Article in Molecular & cellular proteomics : MCP, 2008. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 11 papers.
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Who cites it
11 citing papers in PubMed, 34 citations in OpenAlex.
- Ultrastructural Study and Immunohistochemical Characteristics of Mesencephalic Tegmentum in Juvenile Chum Salmon (International journal of molecular sciences · 2025Article
- Identification of TSPAN4 as Novel Histamine HBiomolecules · 2021Article
- Melatonin receptors: molecular pharmacology and signalling in the context of system bias.British journal of pharmacology · 2018Review
- Melatonin and Hippo Pathway: Is There Existing Cross-Talk?International journal of molecular sciences · 2017Article
- Systematic protein-protein interaction mapping for clinically relevant human GPCRs.Molecular systems biology · 2017Article
- Update on melatonin receptors: IUPHAR Review 20.British journal of pharmacology · 2016Review
- Melatonin and pancreatic islets: interrelationships between melatonin, insulin and glucagon.International journal of molecular sciences · 2013Review
- Identification of GPCR-interacting cytosolic proteins using HDL particles and mass spectrometry-based proteomic approach.PloS one · 2013Article
- Molecular organization and dynamics of the melatonin MT₁ receptor/RGS20/G(i) protein complex reveal asymmetry of receptor dimers for RGS and G(i) coupling.The EMBO journal · 2010Article
- Ligand-based peptide design and combinatorial peptide libraries to target G protein-coupled receptors.Current pharmaceutical design · 2010Review
- Melatonin receptors, heterodimerization, signal transduction and binding sites: what's new?British journal of pharmacology · 2008Review
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Authors and funding
13 authors at 6 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
G protein-coupled receptors (GPCRs) constitute the largest family of membrane receptors and are major drug targets. Recent progress has shown that GPCRs are part of large protein complexes that regulate their activity. We present here a generic approach for identification of these complexes that is based on the use of receptor subdomains and that overcomes the limitations of currently used genetics and proteomics approaches. Our approach consists of a carefully balanced combination of chemically synthesized His6-tagged baits, immobilized metal affinity chromatography, one- and two-dimensional gel electrophoresis separation and mass spectrometric identification. The carboxyl-terminal tails (C-tails) of the human MT1 and MT2 melatonin receptors, two class A GPCRs, were used as models to purify protein complexes from mouse brain lysates. We identified 32 proteins that interacted with the C-tail of MT1, 14 proteins that interacted with the C-tail of MT2, and eight proteins that interacted with both C-tails. Several randomly selected proteins were validated by Western blotting, and the functional relevance of our data was further confirmed by showing the interaction between the full-length MT1 and the regulator of G protein signaling Z1 in transfected HEK 293 cells and native tissue. Taken together, we have established an integrated and generic purification strategy for the identification of high quality and functionally relevant GPCR-associated protein complexes that significantly widens the repertoire of available techniques.
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