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Cationic antimicrobial (host defense) peptides (CAPs) are able to kill microorganisms and cancer cells, leading to their consideration as novel candidate therapeutic agents in human medicine. CAPs can physically associate with anionic membrane structures, such as those found on c ...

Changes in proteolytic activity are associated with several diseases, including cancer. Proteases are potential drug targets and targeting of proteases is used for treatment of various conditions/diseases, like high blood pressure and HIV. We present here detailed protocols for b ...

Angiotensin II (AngII) is an octapeptide hormone with a key role in blood pressure regulation. AngII increases blood pressure by stimulating G protein-coupled receptors in vascular smooth muscle. AngII receptors are therefore an important target in patients with high blood pressure. ...

Expression of proteins in insect cells using recombinant baculoviruses has gained wide use in the G protein-coupled receptor (GPCR) community. This expression system produces high yields of functional receptor, is able to perform post-translational modifications, and is readily ...

Over 1000 sequences likely to encode G protein-coupled receptors (GPCRs) are currently available in publicly accessible and proprietary databases and this number may grow with the refinement of a number of different genomes. However, despite recent efforts in the crystallization of t ...

An overview of the characteristics of classical and outlier-resistant data summaries is provided. The latter are important because outlier data can skew results and decisions based on them. The simple data summaries are the basis for all composite assay and screening data quality measur ...

Understanding the ternary complex between G protein-coupled receptors (GPCRs), cognate G proteins, and their ligands is an important landmark for drug discovery. Yet, little is known about the specific interactions between GPCRs and G proteins. For a better perspective on the ternary co ...

G protein-coupled receptors (GPCRs) represent 50–60% of the current drug targets. There is no doubt that this family of membrane proteins plays a crucial role in drug discovery today. Classically, a number of drugs based on GPCRs have been developed for such different indications as cardiovas ...

Modulators of G protein-coupled receptors (GPCRs) form a key area for the pharmaceutical industry, representing ˜27% of all Food and Drug Administration (FDA)-approved drugs. Consequently, there are a wide variety of in vitro plate-based screening technologies that enable the measu ...

The discovery of drugs for G protein-coupled receptors (GPCRs) has traditionally been very successful, even before the structural nature of these molecular targets was elucidated. Over the years, this family of proteins has become more important in the understanding and treatment of di ...

G protein-coupled receptors (GPCRs) are integral membrane proteins with seven transmembrane-spanning α-helices. Following ligand activation, many GPCRs are rapidly phosphorylated on serine/threonine residues in their cytoplasmic domains, principally the carboxyl ...

Deorphanization of the large group of G protein-coupled receptors (GPCRs) for which an endogenous activating ligand has not yet been identified (orphan GPCRs) has become increasingly difficult. A specialized technique that has been successfully applied to deorphanize some of these ...

A robust method to directly measure ligand–receptor binding interactions using fluorescence cross-correlation spectroscopy (FCCS) is described. The example receptor systems demonstrated here are the human mu-opioid receptor, a representative G protein-coupled recep ...

The measurement of cyclic adenosine monophosphate levels through reporter gene technology represents one of the most popular and cost-effective methods to assess changes in functional activity of G protein-coupled receptors (GPCRs). This chapter provides a generic protocol for ...

Bioluminescence resonance energy transfer (BRET) is a powerful and increasingly popular technique for studying protein–protein interactions in live cells and real time. In particular, there has been considerable interest in the ability to monitor interactions between G protei ...

Fluorescence resonance energy transfer (FRET) has been utilized to determine distances between a fluorescence donor and a fluorescence acceptor having appropriately overlapping spectra. In this chapter, we utilize this approach to establish distances between a fluorescen ...

Fluorescence techniques can provide insights into the environment of fluorescent indicators incorporated within a ligand as it is bound to its receptor. Fluorescent indicators of different sizes and chemical characteristics ranging from hydrophilic to hydrophobic that are i ...

G protein-coupled receptors (GPCRs) are a large family of proteins that represent targets for approximately 40% of all approved drugs. They possess unique structural motifs that allow them to interact with a diverse series of extracellular ligands, as well as intracellular signaling pr ...

Fluorescent dyes sensitive to changes in intracellular calcium have become increasingly popular in G protein-coupled receptor (GPCR) drug discovery for several reasons. First of all, the assays using the dyes are easy to perform and are of low cost compared to other assays. Second, most non-G ...

The C-termini of G protein-coupled receptors (GPCRs) interact with specific kinases and arrestins in an agonist-dependent manner suggesting that conformational changes induced by ligand binding within the transmembrane domains are transmitted to the C-terminus. F�rster res ...