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Related Concept Videos

GPCR Desensitization01:12

GPCR Desensitization

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G protein-coupled receptor (GPCR) signaling plays a crucial role in cell functioning. GPCR desensitization is an equally essential process. It allows cells to respond to changing environments and regain sensitivity to new stimuli while preventing unnecessary stimulation when no longer needed. Prolonged exposure to stimuli leads to GPCR desensitization. It involves blocking the receptors from binding and activating additional G proteins. This inhibits activation of downstream effectors, thereby...
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Receptor-mediated Endocytosis01:20

Receptor-mediated Endocytosis

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Receptor-mediated endocytosis is when bulk amounts of specific molecules are imported into a cell after binding to cell surface receptors. The molecules bound to these receptors are taken into the cell through inward folding of the cell surface membrane, which is eventually pinched off into a vesicle within the cell. Structural proteins, such as clathrin, coat the budding vesicle.
Clathrin-Mediated Endocytosis of LDL
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GPI Anchoring of Proteins in the ER Membrane01:29

GPI Anchoring of Proteins in the ER Membrane

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GPI-anchoring is a post-translational, reversible protein modification that is ubiquitous in eukaryotes. Such proteins are primarily present on the exoplasmic leaflet of the plasma membrane.
GPI-anchor structure
A sequence of 11 enzymatic reactions results in the synthesis of the complete GPI anchor consisting of a hydrophobic and a hydrophilic portion. The hydrophobic portion comprises phosphatidylinositol, while the hydrophilic part comprises polar groups like phosphoethanolamine,...
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Directing Proteins to the Rough Endoplasmic Reticulum01:34

Directing Proteins to the Rough Endoplasmic Reticulum

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The organelle-specific signaling sequences direct proteins synthesized in the cytosol to their final destination like ER, mitochondria, peroxisomes, etc. Some of the proteins directed to ER are then trafficked via vesicles to other organelles within the cell or the extracellular environment through the Golgi complex. For example, the rough ER synthesizes soluble proteins for transportation to the lysosomes or secretion out of the cell. It can also synthesize transmembrane proteins that can...
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The Early Endosome: Endocytosis of Transferrin01:28

The Early Endosome: Endocytosis of Transferrin

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Essential proteins such as insulin or low-density lipoprotein (LDL) and micronutrients such as iron enter a eukaryotic cell through receptor-mediated endocytosis. Subsequently, the early endosomes fuse with the vesicles containing such receptor-ligand complexes and play a vital role in sorting the incoming ligands and receptors. While the ligands are either degraded inside the vesicle or released into the cytosol, their receptors are returned to the plasma membrane for further rounds of...
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Receptor Downregulation in MVBs01:15

Receptor Downregulation in MVBs

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Multivesicular bodies (MVBs) are mature endosomes that sort ubiquitinated proteins and then fuse with lysosomes to degrade the sorted proteins. Epidermal growth factor (EGF) and its receptor (EGFR) form a complex that can be internalized through endocytosis, sorted into an MVB, and later degraded.
The EGFR can initiate signaling pathways that  lead to cell proliferation, migration, and differentiation. Overexpression of EGFR  stimulates cells to proliferate. Excessive  EGFR...
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Related Experiment Video

Updated: Jul 21, 2025

Imaging G-protein Coupled Receptor GPCR-mediated Signaling Events that Control Chemotaxis of Dictyostelium Discoideum
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Imaging G-protein Coupled Receptor GPCR-mediated Signaling Events that Control Chemotaxis of Dictyostelium Discoideum

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Endosome positioning coordinates spatially selective GPCR signaling.

Blair K A Willette1, Jin-Fan Zhang2,3, Jin Zhang2,3,4

  • 1Department of Pharmacology and Cancer Biology, Duke University, Durham, NC, USA.

Nature Chemical Biology
|July 27, 2023
PubMed
Summary
This summary is machine-generated.

G-protein-coupled receptors (GPCRs) initiate distinct cellular responses based on their location. This study reveals that the physical positioning of GPCR-containing endosomes, not just their biochemistry, is crucial for compartmentalized signaling.

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Visualizing Clathrin-mediated Endocytosis of G Protein-coupled Receptors at Single-event Resolution via TIRF Microscopy
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Detection of Ligand-activated G Protein-coupled Receptor Internalization by Confocal Microscopy
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Detection of Ligand-activated G Protein-coupled Receptor Internalization by Confocal Microscopy

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Imaging G-protein Coupled Receptor GPCR-mediated Signaling Events that Control Chemotaxis of Dictyostelium Discoideum
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Visualizing Clathrin-mediated Endocytosis of G Protein-coupled Receptors at Single-event Resolution via TIRF Microscopy
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Detection of Ligand-activated G Protein-coupled Receptor Internalization by Confocal Microscopy
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Detection of Ligand-activated G Protein-coupled Receptor Internalization by Confocal Microscopy

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Area of Science:

  • Cellular Biology
  • Molecular Pharmacology
  • Biophysics

Background:

  • G-protein-coupled receptors (GPCRs) exhibit functional diversity based on their subcellular localization.
  • Previous research focused on biochemical regulation of compartmentalized GPCR signaling.
  • The role of physical receptor positioning in GPCR signaling remains underexplored.

Purpose of the Study:

  • To investigate the biophysical positioning of receptor-containing endosomes as a mechanism for compartmentalized GPCR signaling.
  • To develop methods for manipulating endosome positioning and measuring downstream signaling.
  • To elucidate the role of endosome localization in GPCR-mediated transcriptional responses.

Main Methods:

  • Developed a method to rapidly and selectively redistribute receptor-containing endosomes in intact cells without altering biochemical composition.
  • Utilized two complementary optical readouts for single-cell resolution measurements of GPCR/cyclic AMP (cAMP)-dependent transcriptional signaling.
  • Assessed bulk and gene-specific signaling responses.

Main Results:

  • Disruption of native endosome positioning significantly inhibited endosome-dependent transcriptional responses.
  • Demonstrated that endosome localization is a key factor in initiating spatially selective GPCR signaling.
  • Identified a mechanistic role for phosphodiesterase (PDE)-mediated cAMP hydrolysis and local protein kinase A (PKA) activity.

Conclusions:

  • Endosome positioning is a principal mediator of spatially selective GPCR signaling.
  • Biophysical localization of receptor-containing endosomes represents a novel regulatory mechanism for GPCR function.
  • This finding offers a new perspective beyond biochemical regulation in understanding compartmentalized GPCR signaling.