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Related Concept Videos

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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Channel Rhodopsins01:11

Channel Rhodopsins

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Most organisms use photoreceptors to sense and respond to light. Examples of photoreceptors include bacteriorhodopsins and bacteriophytochromes in some bacteria, phytochromes in plants, and rhodopsins in the photoreceptor cells of the vertebral retina. The light-sensitive property of these receptors is because of the bound chromophores, such as bilin in the phytochromes and retinal in the rhodopsins.
Rhodopsins belong to the family of cell surface proteins called G-protein coupled receptors,...
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G-Protein Gated Ion Channels01:21

G-Protein Gated Ion Channels

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GPCRs are primarily responsible for our sense of smell, taste, and vision.  The binding of a sensory stimulus activates GPCR to stimulate effector proteins, many of which are ion channels in the sensory organs. GPCRs modulate the opening and closing of the target ion channels either directly by binding them, or by releasing second messengers that activate these channels. As ions move across the membrane, the membrane potential is altered, which induces an appropriate response.
Sensory...
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G Protein-coupled Receptors01:15

G Protein-coupled Receptors

12.1K
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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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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Transducer Mechanism: G Protein–Coupled Receptors01:30

Transducer Mechanism: G Protein–Coupled Receptors

2.0K
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.
GPCRs are also called heptahelical,...
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Related Experiment Video

Updated: Jul 4, 2025

Strategic Screening and Characterization of the Visual GPCR-mini-G Protein Signaling Complex for Successful Crystallization
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Strategic Screening and Characterization of the Visual GPCR-mini-G Protein Signaling Complex for Successful Crystallization

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Scaling up Functional Analyses of the G Protein-Coupled Receptor Rhodopsin.

Benjamin M Scott1, Steven K Chen1, Alexander Van Nynatten1

  • 1Department of Cell and Systems Biology, University of Toronto, Toronto, ON, Canada.

Journal of Molecular Evolution
|February 7, 2024
PubMed
Summary
This summary is machine-generated.

We developed a high-throughput assay to measure mutations in rhodopsin, a G protein-coupled receptor (GPCR). This deep mutational scanning method reveals how mutations affect rhodopsin activation and protein structure, aiding disease variant understanding.

Keywords:
G protein-coupled receptorHigh-throughput yeast assayRhodopsin structure and function

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G Protein-selective GPCR Conformations Measured Using FRET Sensors in a Live Cell Suspension Fluorometer Assay
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A Rhodopsin Transport Assay by High-Content Imaging Analysis
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A Rhodopsin Transport Assay by High-Content Imaging Analysis

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Related Experiment Videos

Last Updated: Jul 4, 2025

Strategic Screening and Characterization of the Visual GPCR-mini-G Protein Signaling Complex for Successful Crystallization
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G Protein-selective GPCR Conformations Measured Using FRET Sensors in a Live Cell Suspension Fluorometer Assay
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A Rhodopsin Transport Assay by High-Content Imaging Analysis
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A Rhodopsin Transport Assay by High-Content Imaging Analysis

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Area of Science:

  • Biochemistry
  • Molecular Biology
  • Genetics

Background:

  • G protein-coupled receptors (GPCRs) mediate cellular responses to external stimuli.
  • Understanding how mutations in GPCR genes impact receptor activation and signaling is crucial.
  • Existing methods for studying GPCR mutations, like deep mutational scanning, lack high-throughput measurement for rhodopsin activation.

Purpose of the Study:

  • To develop and scale a high-throughput fluorescent reporter assay in budding yeast for measuring rhodopsin activation.
  • To investigate the functional effects of over 1200 individual human rhodopsin mutants.
  • To analyze mutational tolerance in rhodopsin based on its 3D structure and correlate findings with disease variants.

Main Methods:

  • Engineered a fluorescent reporter assay in budding yeast to study rhodopsin's light-activated signal transduction.
  • Generated over 1200 human rhodopsin mutants using low-frequency random mutagenesis of the RHO gene.
  • Performed deep mutational scanning to measure the functional impact of each mutant.

Main Results:

  • Identified over 1200 rhodopsin mutants and measured their functional effects on activation.
  • Determined that transmembrane helices are less mutation-tolerant than helices facing the lipid bilayer, indicating structure-dependent mutational tolerance.
  • Found that many pathogenic rhodopsin mutants exhibit loss-of-function, consistent with clinical observations and a complex counterion mechanism.

Conclusions:

  • Deep mutational scanning is a viable and effective method for studying rhodopsin activation.
  • The study provides insights into mutational tolerance within rhodopsin structure, potentially applicable to other transmembrane proteins.
  • Findings contribute to understanding rhodopsin-related diseases and the mechanisms of receptor activation.