Next Article in Journal
Evaluation of a Method for Nitrotyrosine Site Identification and Relative Quantitation Using a Stable Isotope-Labeled Nitrated Spike-In Standard and High Resolution Fourier Transform MS and MS/MS Analysis
Next Article in Special Issue
Alpha-Bulges in G Protein-Coupled Receptors
Previous Article in Journal
Preparation of Lung-Targeting, Emodin-Loaded Polylactic Acid Microspheres and Their Properties
Previous Article in Special Issue
Differential Signaling by Protease-Activated Receptors: Implications for Therapeutic Targeting
Article Menu

Export Article

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

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

1
Department of Pharmacy, University of Pisa, 56126 Pisa, Italy
2
Department of Pharmacological and Biomolecular Sciences, University of Milan, 20133 Milan, Italy
*
Author to whom correspondence should be addressed.
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)
View Full-Text   |   Download PDF [719 KB, uploaded 19 June 2014]   |  

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 immunoenzymatic assay; G protein coupled-receptors; protein-protein interactions
Figures

This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).

Scifeed alert for new publications

Never miss any articles matching your research from any publisher
  • Get alerts for new papers matching your research
  • Find out the new papers from selected authors
  • Updated daily for 49'000+ journals and 6000+ publishers
  • Define your Scifeed now

SciFeed Share & Cite This Article

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.

Show more citation formats Show less citations formats

Related Articles

Article Metrics

Article Access Statistics

1

Comments

[Return to top]
Int. J. Mol. Sci. EISSN 1422-0067 Published by MDPI AG, Basel, Switzerland RSS E-Mail Table of Contents Alert
Back to Top