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Related Concept Videos

Clathrin Coated Vesicles01:12

Clathrin Coated Vesicles

Clathrin-coated vesicles use endocytosis to transport receptors and lysosomal hydrolases from the Golgi to the lysosome in the late secretory pathway. Clathrin-mediated endocytosis was the first described endocytic process, and Clathrin-coated vesicles remain one of the most well-studied transport vesicles. The molecular machinery that generates clathrin-coated vesicles comprises over 50 proteins that precisely coordinate vesicle formation. Cell surface receptors concentrated in indented sites...
Receptor Downregulation in MVBs01:15

Receptor Downregulation in MVBs

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 activation may...
Regulation of Nuclear Protein Sorting01:45

Regulation of Nuclear Protein Sorting

Nuclear protein sorting regulates nucleus composition and gene expression, crucial for determining the fate of a eukaryotic cell. Hence, the entry and exit of molecules across the nuclear envelope is a tightly controlled process. Nuclear protein sorting can be inhibited by one of the following ways: 1) masking cargo signal sequences, 2) modifying the nuclear receptor's affinity for cargo, 3) controlling the nuclear pore size, 4) retaining the cargo during its transit to the cytosol or the...
Membrane Asymmetry Regulating Transporters01:19

Membrane Asymmetry Regulating Transporters

Enzymes like flippase, floppase, and scramblase transfer phospholipids from one layer to another in the membrane, thereby affecting membrane asymmetry.
Flippase
Eukaryotic flippases are type-IV P-type ATPases or P4-ATPases belonging to P-type ATPase family proteins that are membrane-bound pumps involved in the ATP-mediated transport of ions and molecules across the membrane. Flippases flip specific phospholipids from the outer to the inner leaflet of a membrane. All P4-ATPases have one...
Pinching-off of Coated Vesicles01:32

Pinching-off of Coated Vesicles

Vesicle budding is orchestrated by distinct cytosolic proteins such as adaptor proteins, coat proteins, and GTPases. To initiate vesicle budding, membrane-bending proteins containing crescent-shaped BAR domains bind to the lipid heads in the bilayer and distort the membrane to form a protein-coated vesicle bud. Adaptors proteins such as AP2 for clathrin-coated vesicles can nucleate on the deformed membrane. Finally, coat proteins such as clathrin or COPI and COPII assemble into a coat forming...
Directing Proteins to the Rough Endoplasmic Reticulum01:34

Directing Proteins to the Rough Endoplasmic Reticulum

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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Related Experiment Video

Updated: Jun 25, 2026

In vivo and in vitro Studies of Adaptor-clathrin Interaction
17:14

In vivo and in vitro Studies of Adaptor-clathrin Interaction

Published on: January 26, 2011

Differential clathrin binding and subcellular localization of OCRL1 splice isoforms.

Rawshan Choudhury1, Christopher J Noakes, Edward McKenzie

  • 1Faculty of Life Sciences, University of Manchester, The Michael Smith Building, Oxford Road, Manchester M13 9PT, United Kingdom.

The Journal of Biological Chemistry
|February 13, 2009
PubMed
Summary

The inositol polyphosphate 5-phosphatase OCRL1 has two isoforms that affect clathrin binding differently. Isoform a binds clathrin more strongly, impacting endocytosis, suggesting distinct functional roles in cellular trafficking.

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SorLA and CLC:CLF-1-dependent Downregulation of CNTFR&#945; as Demonstrated by Western Blotting, Inhibition of Lysosomal Enzymes, and Immunocytochemistry
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SorLA and CLC:CLF-1-dependent Downregulation of CNTFRα as Demonstrated by Western Blotting, Inhibition of Lysosomal Enzymes, and Immunocytochemistry

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Visualizing Clathrin-mediated Endocytosis of G Protein-coupled Receptors at Single-event Resolution via TIRF Microscopy
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Visualizing Clathrin-mediated Endocytosis of G Protein-coupled Receptors at Single-event Resolution via TIRF Microscopy

Published on: October 20, 2014

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In vivo and in vitro Studies of Adaptor-clathrin Interaction
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Published on: January 26, 2011

SorLA and CLC:CLF-1-dependent Downregulation of CNTFR&#945; as Demonstrated by Western Blotting, Inhibition of Lysosomal Enzymes, and Immunocytochemistry
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SorLA and CLC:CLF-1-dependent Downregulation of CNTFRα as Demonstrated by Western Blotting, Inhibition of Lysosomal Enzymes, and Immunocytochemistry

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Visualizing Clathrin-mediated Endocytosis of G Protein-coupled Receptors at Single-event Resolution via TIRF Microscopy
12:40

Visualizing Clathrin-mediated Endocytosis of G Protein-coupled Receptors at Single-event Resolution via TIRF Microscopy

Published on: October 20, 2014

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Genetics

Background:

  • OCRL1 mutations cause Lowe syndrome, affecting brain, kidney, and eye development.
  • OCRL1 has two splice isoforms (a and b) differing by 8 amino acids.
  • The functional significance of OCRL1 splicing and its impact on protein interactions remain unclear.

Purpose of the Study:

  • To investigate the impact of OCRL1 splicing on its interaction with clathrin.
  • To determine how OCRL1 isoforms differ in their association with clathrin-coated vesicles.
  • To elucidate the role of OCRL1 in clathrin-mediated endocytosis.

Main Methods:

  • Comparative analysis of clathrin binding affinity between OCRL1 isoforms a and b.
  • Immunofluorescence and biochemical assays to detect enrichment in clathrin-coated trafficking intermediates.
  • Functional assays examining transferrin endocytosis in cells expressing OCRL1 variants.
  • Investigation of OCRL1 interaction with Rab GTPases and AP2.

Main Results:

  • OCRL1 isoform a exhibits higher affinity for clathrin compared to isoform b.
  • Isoform a is significantly more enriched in clathrin-coated trafficking intermediates.
  • A second clathrin-binding site was identified in OCRL1.
  • OCRL1 association with clathrin-coated intermediates depends on membrane association via Rab GTPases, not AP2.
  • Expression of OCRL1 isoform a lacking the 5-phosphatase domain impairs transferrin endocytosis, unlike isoform b.

Conclusions:

  • OCRL1 splicing generates two functional pools with distinct roles in clathrin-mediated trafficking.
  • Isoform a plays a significant role in clathrin-mediated endocytosis, while isoform b has a lesser or no role.
  • These findings provide insights into the molecular mechanisms underlying Lowe syndrome and cellular trafficking pathways.