Characterization of the interaction between the prostaglandin D2 DP1 receptor and the intracellular L-prostaglandin D synthase

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 1234; p. 53
• Main Authors: Binda, Chantal, Parent, Jean-Luc
• Format: Journal Article
• Language: English
• Published: United States 2015
• Subjects: Cell Culture Techniques; Cell Line; Humans; Immunoprecipitation - methods; Intracellular Space - metabolism; Intramolecular Oxidoreductases - genetics; Intramolecular Oxidoreductases - metabolism; Lipocalins - genetics; Lipocalins - metabolism; Protein Binding; Receptors, Immunologic - genetics; Receptors, Immunologic - metabolism; Receptors, Prostaglandin - genetics; Receptors, Prostaglandin - metabolism; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism
• DOI: 10.1007/978-1-4939-1755-6_6

Identification of G protein-coupled receptor (GPCR)-interacting proteins is an intense subject of current research. However, confirmation and characterization of identified interactions can be difficult with GPCRs, especially at the endogenous level. Here, we describe how we characterized the interaction between the prostaglandin D2 DP1 receptor and the intracellular L-type prostaglandin D synthase by in vitro pull-down assays using purified recombinant GST- and His-tagged proteins, by co-immunoprecipitation of overexpressed Flag- and HA-tagged proteins, and by co-immunoprecipitation of endogenous proteins.