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Related Concept Videos

G-protein Coupled Receptors01:21

G-protein Coupled Receptors

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G-protein coupled receptors are ligand binding receptors that indirectly affect changes in the cell. The actual receptor is a single polypeptide that transverses the cell membrane seven times creating intracellular and extracellular loops. The extracellular loops create a ligand specific pocket which binds to neurotransmitters or hormones. The intracellular loops holds onto the G-protein.
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G Protein-coupled Receptors01:15

G Protein-coupled Receptors

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G Protein-Coupled Receptors or GPCRs are membrane-bound receptors that transiently associate with heterotrimeric G proteins and induce an appropriate response to sensory stimuli such as light, odors, hormones, cytokines, or neurotransmitters.
GPCRs are also called heptahelical, 7TM, or serpentine receptors, and consist of seven (H1-H7) transmembrane alpha-helices that span the bilayer to form a cylindrical core. The transmembrane helices are connected by three extracellular loops and three...
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Activation and Inactivation of G Proteins01:22

Activation and Inactivation of G Proteins

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Heterotrimeric G proteins are guanine nucleotide-binding proteins. As the name suggests, heterotrimeric G proteins are composed of three subunits: alpha, beta, and gamma. They remain GDP-bound or GTP-bound inside the cells and switch between inactive/active states. The Gα subunit possesses the nucleotide-binding pocket that binds guanine nucleotides and switches between GDP or GTP-bound states. In contrast, the Gꞵ and Gγ subunits are always bound together with high...
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Updated: Dec 3, 2025

Genetically-encoded Molecular Probes to Study G Protein-coupled Receptors
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G-Protein-Coupled Receptor Expression and Purification.

Karolina Corin1, Lotta T Tegler2, Sotirios Koutsopoulos3

  • 1Department of Chemistry and Biochemistry, UCLA-DOE Institute, Molecular Biology Institute, University of California, Los Angeles, CA, USA.

Methods in Molecular Biology (Clifton, N.J.)
|October 31, 2020
PubMed
Summary

Producing stable G-protein-coupled receptors (GPCRs) is crucial for research and drug development. This study presents three effective protocols for GPCR production and solubilization using cell-free systems, HEK cells, and E. coli.

Keywords:
DetergentsG-protein-coupled receptor (GPCR )Olfactory receptors, membrane proteinsSurfactants

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Area of Science:

  • Membrane protein biochemistry
  • Structural biology
  • Drug discovery

Background:

  • G-protein-coupled receptors (GPCRs) are vital cell membrane proteins regulating numerous biological functions and serving as drug targets.
  • Limited understanding of GPCR structure and function hinders research and therapeutic development.
  • A primary challenge is producing sufficient quantities of soluble, functional, and stable GPCRs for study.

Purpose of the Study:

  • To present three laboratory protocols for producing and solubilizing stable GPCRs.
  • To address the bottleneck in GPCR production for enhanced structural and functional analysis.
  • To facilitate GPCR research and drug targeting initiatives.

Main Methods:

  • Developed and employed three distinct production systems: cell-free in vitro translation, HEK cells, and Escherichia coli.
  • Utilized specific detergents for GPCR extraction and stabilization outside the cell membrane.
  • Purified stable receptors via immunoaffinity chromatography and gel filtration.

Main Results:

  • Successfully produced and solubilized stable GPCRs using the three described methods.
  • Demonstrated the utility of immunoaffinity chromatography and gel filtration for purification.
  • Confirmed the applicability of standard biophysical techniques and biochemical assays for analyzing purified receptors.

Conclusions:

  • The presented protocols offer effective strategies for obtaining stable, soluble GPCRs.
  • These methods overcome key production challenges, enabling further structural and functional investigations.
  • Facilitates advancements in GPCR research and the development of novel therapeutics targeting these important proteins.