How is GPCR regulated?

Every animal species expresses hundreds of different G protein-coupled receptors (GPCRs) that respond to a wide variety of external stimuli. GPCRs-driven signaling pathways are involved in pretty much every physiological function and in many pathologies.

What is GPCR pharmacology?

GPCRs are 7-transmembrane integral membrane proteins that typically translate extracellular stimulation into intracellular signals. GPCR activation is usually mediated by agonist binding which stabilizes receptor conformations that recruit and ultimately activate intracellular transducers.

How many types of GPCR are there?

There are numerous different types of GPCRs—some 1,000 types are encoded by the human genome alone—and as a group they respond to a diverse range of substances, including light, hormones, amines, neurotransmitters, and lipids.

What is a Class B GPCR?

Structurally, class B GPCRs consist of a large N-terminal extracellular domain (ECD) and a transmembrane domain (TMD) comprising the ‘GPCR’ signature of seven membrane spanning α-helices, which is involved in signalling via coupling to heterotrimeric G proteins that primarily activate adenylate cyclase to increase the …

What are B Arrestins?

beta-Arrestins are versatile adapter proteins that form complexes with most G-protein-coupled receptors (GPCRs) following agonist binding and phosphorylation of receptors by G-protein-coupled receptor kinases (GRKs).

What is Htrf?

HTRF (Homogeneous Time Resolved Fluorescence) is the most frequently used generic assay technology to measure analytes in a homogenous format, which is the ideal platform used for drug target studies in high-throughput screening (HTS).

How are cAMP levels measured?

The cAMP-Glo™ Max Assay measures cAMP levels through protein kinase A (PKA), which is activated upon release of its regulatory subunits after binding to cAMP (Figure 1). PKA is a component of the cAMP Detection Solution, and once activated, PKA will use any ATP present to phosphorylate its substrate.

What is the structure of GPCR?

In terms of structure, GPCRs are characterized by an extracellular N-terminus, followed by seven transmembrane (7-TM) α-helices (TM-1 to TM-7) connected by three intracellular (IL-1 to IL-3) and three extracellular loops (EL-1 to EL-3), and finally an intracellular C-terminus.

How are GPCR signaling assays used in medicine?

GPCR Signaling Assays G-protein-coupled receptors (GPCR) represent one of the most important classes of drug targets. We offer a range of easy-to-use, bioluminescence-based GPCR-related assays, ranging from assays that can be used to measure cAMP levels in cells to kits that measure GTPase and PDE activity. Learn about GPCR Signaling Assays

Where can I buy pathhunter arrestin GPCR assays?

DiscoveRx Corporation 42501 Albrae Street Fremont, CA 94538 USA tel | 1.510.979.1415 x104 [email protected] For some products/cell lines, certain 3rdparty gene specific patents may be required to use the cell line. It is the purchaser’s responsibility to determine if such patents or other intellectual property rights are required.

Which is the enzyme acceptor for pathhunter GPCR?

This PathHunter system combines engineered clonal cell lines stably expressing the ProLink™ (PK)- tagged GPCR of interest and the Enzyme acceptor (EA)-tagged β-Arrestin fusion proteins with optimized PathHunter Detection Reagents (Cat. #93-0001, 93-0001L and 93-0001XL).

How is GPCR fused with a β-galactosidase?

The GPCR is fused in frame with a small enzyme donor fragment ProLink™ (PK) and co-expressed in cells stably expressing a fusion protein of β-arrestin and the larger, N-terminal deletion mutant of β-galactosidase (called enzyme acceptor or EA).

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