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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

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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.
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Transducer Mechanism: G Protein–Coupled Receptors01:30

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G Protein–Coupled Receptors (GPCRs) are membrane-bound receptors that transiently associate with heterotrimeric G proteins and induce an appropriate response to various stimuli. GPCRs regulate critical physiological pathways and are excellent drug targets for treating diseases such as diabetes, cancer, obesity, depression, or Alzheimer's. Nearly 35% of approved drugs implement their therapeutic effects by selectively interacting with specific GPCRs.
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Habitat Fragmentation02:31

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Habitat fragmentation describes the division of a more extensive, continuous habitat into smaller, discontinuous areas. Human activities such as land conversion, as well as slower geological processes leading to changes in the physical environment, are the two leading causes of habitat fragmentation. The fragmentation process typically follows the same steps: perforation, dissection, fragmentation, shrinkage, and attrition.
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Electron Affinity03:07

Electron Affinity

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The electron affinity (EA) is the energy change for adding an electron to a gaseous atom to form an anion (negative ion).
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Related Experiment Video

Updated: Feb 8, 2026

Characterization of G Protein-coupled Receptors by a Fluorescence-based Calcium Mobilization Assay
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Characterization of G Protein-coupled Receptors by a Fluorescence-based Calcium Mobilization Assay

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Simplifying G Protein-Coupled Receptor Isolation with a Calcium-Dependent Fragment Complementation Affinity System.

Niamh Ní Mhurchú1, Lioudmila Zoubak2, Gavin McGauran1

  • 1School of Biomolecular and Biomedical Science, Conway Institute , University College Dublin , Dublin D04 V1W8 , Ireland.

Biochemistry
|June 30, 2018
PubMed
Summary
This summary is machine-generated.

Researchers developed a novel molecular switch system for capturing and purifying G protein-coupled receptors. This method simplifies purification, yielding functional proteins in a simple buffer, unlike traditional complex protocols.

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Protein Purification

Background:

  • Isolating recombinant G protein-coupled receptors (GPCRs) from membrane preparations is complex, requiring harsh solubilization conditions and multi-step affinity chromatography.
  • Detergent micelles, high salt concentrations, and glycerol stabilize GPCRs but impede standard affinity resin performance and necessitate complicating elution agents.

Purpose of the Study:

  • To develop a simplified and highly specific affinity chromatography system for the isolation and purification of recombinant membrane-bound proteins, specifically targeting G protein-coupled receptors.
  • To introduce a novel high-affinity fragment complementation molecular switch for efficient receptor capture and a calcium chelation-based elution strategy.

Main Methods:

  • Design and implementation of a high-affinity fragment complementation molecular switch system.
  • Application of the system for the isolation and purification of the cannabinoid receptor 2 (CB2) from membrane preparations.
  • Comparison of the novel system with commercially available affinity chromatography methods.

Main Results:

  • The novel system effectively captures target receptors in stringent solubilization buffers.
  • A calcium chelation-based elution releases functional protein in a simple buffer, simplifying downstream applications.
  • Demonstrated successful isolation and purification of the CB2 receptor, with performance compared to existing commercial systems.

Conclusions:

  • The developed fragment complementation molecular switch offers a powerful and simplified approach for purifying challenging recombinant membrane-bound proteins.
  • This novel affinity chromatography system overcomes limitations of traditional methods, enhancing yield and purity for various protein targets.
  • The system holds broad applicability for studying any recombinant membrane-bound protein, with potential for further optimization.