Recent methodological advances in the discovery of GPCR-associated protein complexes

Protein-interaction networks have important roles in cellular homeostasis and the generation of complexity in biological systems. G-protein-coupled receptors (GPCRs), the largest family of membrane receptors and important drug targets, are integral parts of these networks. Ligand stimulation and the dynamic interaction with GPCR-associated protein complexes (GAPCs) constitute two important regulatory mechanisms of GPCR function. Several genomic and proteomic approaches have been developed to identify GAPCs in the past. However, this task turned out to be particularly demanding owing to difficulties in preserving the complex three-dimensional GPCR structure during receptor solubilization and to inherent limitations in the use of isolated receptor domains as bait. Newly emerging methods have the potential to overcome these limitations and will certainly boost the identification of functionally relevant GAPCs to finally increase our knowledge of the regulation of GPCRs and provide novel drug targets. Here, we focus on the comparison of two complementary GAPC purification strategies, which are based on soluble GPCR subdomains and entire GPCRs.