A Rapid and Efficient Immunoenzymatic Assay to Detect Receptor Protein Interactions: G Protein-Coupled Receptors

Zappelli, Elisa; Daniele, Simona; Abbracchio, Maria P.; Martini, Claudia; Trincavelli, Maria Letizia

doi:10.3390/ijms15046252

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A Rapid and Efficient Immunoenzymatic Assay to Detect Receptor Protein Interactions: G Protein-Coupled Receptors

¹,

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Maria Letizia Trincavelli

Department of Pharmacy, University of Pisa, 56126 Pisa, Italy

Department of Pharmacological and Biomolecular Sciences, University of Milan, 20133 Milan, Italy

Author to whom correspondence should be addressed.

Int. J. Mol. Sci. 2014, 15(4), 6252-6264; https://doi.org/10.3390/ijms15046252

Received: 27 January 2014 / Revised: 10 March 2014 / Accepted: 1 April 2014 / Published: 11 April 2014

(This article belongs to the Collection G Protein-Coupled Receptor Signaling and Regulation)

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Abstract

G protein-coupled receptors (GPCRs) represent one of the largest families of cell surface receptors, and are the target of at least one-third of the current therapeutic drugs on the market. Along their life cycle, GPCRs are accompanied by a range of specialized GPCR-interacting proteins (GIPs), which take part in receptor proper folding, targeting to the appropriate subcellular compartments and in receptor signaling tasks, and also in receptor regulation processes, such as desensitization and internalization. The direction of protein-protein interactions and multi-protein complexes formation is crucial in understanding protein function and their implication in pathological events. Although several methods have been already developed to assay protein complexes, some of them are quite laborious, expensive, and, more important, they do not generate fully quantitative results. Herein, we show a rapid immunoenzymatic assay to quantify GPCR interactionswith its signaling proteins. The recently de-orphanized GPCR, GPR17, was chosen as a GPCR prototype to optimize the assay. In a GPR17 transfected cell line and primary oligodendrocyte precursor cells, GPR17 interaction with proteins involved in the typical GPCR regulation, such as desensitization and internalization machinery, was investigated. The obtained results were validated by co-immunoprecipitation experiments, confirming this new method as a rapid and quantitative assay to study protein-protein interactions. View Full-Text

Keywords: immunoenzymatic assay; G protein coupled-receptors; protein-protein interactions

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MDPI and ACS Style

Zappelli, E.; Daniele, S.; Abbracchio, M.P.; Martini, C.; Trincavelli, M.L. A Rapid and Efficient Immunoenzymatic Assay to Detect Receptor Protein Interactions: G Protein-Coupled Receptors. Int. J. Mol. Sci. 2014, 15, 6252-6264. https://doi.org/10.3390/ijms15046252

AMA Style

Zappelli E, Daniele S, Abbracchio MP, Martini C, Trincavelli ML. A Rapid and Efficient Immunoenzymatic Assay to Detect Receptor Protein Interactions: G Protein-Coupled Receptors. International Journal of Molecular Sciences. 2014; 15(4):6252-6264. https://doi.org/10.3390/ijms15046252

Chicago/Turabian Style

Zappelli, Elisa, Simona Daniele, Maria P. Abbracchio, Claudia Martini, and Maria L. Trincavelli. 2014. "A Rapid and Efficient Immunoenzymatic Assay to Detect Receptor Protein Interactions: G Protein-Coupled Receptors" International Journal of Molecular Sciences 15, no. 4: 6252-6264. https://doi.org/10.3390/ijms15046252

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A Rapid and Efficient Immunoenzymatic Assay to Detect Receptor Protein Interactions: G Protein-Coupled Receptors

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