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  • Title: Cell imaging assays for G protein-coupled receptor internalization: application to high-throughput screening.
    Author: Lee S, Howell B, Kunapuli P.
    Journal: Methods Enzymol; 2006; 414():79-98. PubMed ID: 17110188.
    Abstract:
    There are a number of assays currently available to study G protein-coupled receptors (GPCRs), including ligand binding and functional assays. The latter category, albeit more complex, offers some obvious advantages over traditional ligand-binding assays. Functional cell-based assays typically include second messenger and reporter gene assays, which depend directly or indirectly on the cellular signaling cascade initiated upon receptor activation, respectively. More recently, cell imaging assays monitoring receptor trafficking are becoming increasingly popular. These assays, described in greater detail in this chapter, are independent of receptor signaling and are thus ideally suited for orphan receptors. In addition, these assays provide a valuable measure of receptor desensitization, an important feature for the use of GPCR agonists as potential therapeutic agents. The most popular GPCR imaging assays are based on the principles of receptor desensitization and internalization monitored directly or indirectly by green fluorescent protein.
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