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Related Concept Videos

Clathrin Coated Vesicles01:12

Clathrin Coated Vesicles

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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...
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Pinching-off of Coated Vesicles01:32

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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...
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COP Coated Vesicles00:59

COP Coated Vesicles

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Membrane-enclosed structures called vesicles transport proteins and lipids across the cell. The vesicles derive their cargo from the plasma membrane, Golgi, ER, or endosome. Coated vesicles are spherical, protein-coated carriers with a 50–100 nm diameter that mediate bidirectional transport between the ER and the Golgi. The distribution of proteins between the ER and Golgi complex is dynamic and is maintained by different coated vesicles. Their formation is driven by the assembly of...
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Coat Assembly and GTPases01:33

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Vesicles incorporate different coat protein subunits in different cell locations, which changes the properties of the coat, such as the shape and geometry of the transport vesicles. Thus, vesicle coat proteins also play a significant role in cargo selection.
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Receptor-mediated Endocytosis01:38

Receptor-mediated Endocytosis

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Overview
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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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Related Experiment Video

Updated: May 6, 2026

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

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Quantifying the dynamic interactions between a clathrin-coated pit and cargo molecules.

Aubrey V Weigel1, Michael M Tamkun, Diego Krapf

  • 1School of Biomedical Engineering and Departments of Biomedical Sciences, Biochemistry and Molecular Biology, and Electrical and Computer Engineering, Colorado State University, Fort Collins, CO 80523.

Proceedings of the National Academy of Sciences of the United States of America
|November 13, 2013
PubMed
Summary
This summary is machine-generated.

Cargo molecules frequently escape clathrin-coated pits (CCPs) before internalization, with binding times much shorter than the endocytic process. This reveals a key source of CCP dynamic heterogeneity in cells.

Keywords:
Levy statisticsTIRFlive-cell imagingsingle-molecule tracking

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In vivo and in vitro Studies of Adaptor-clathrin Interaction
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Characterizing the Composition of Molecular Motors on Moving Axonal Cargo Using "Cargo Mapping" Analysis
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In vivo and in vitro Studies of Adaptor-clathrin Interaction
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Area of Science:

  • Cell biology
  • Molecular dynamics
  • Membrane trafficking

Background:

  • Clathrin-mediated endocytosis (CME) is crucial for cellular processes, involving cargo recruitment into clathrin-coated pits (CCPs).
  • Understanding cargo-CCP interaction dynamics is limited due to CCP lifetime heterogeneity and time-dependent interactions.

Purpose of the Study:

  • To directly visualize and quantify the dynamic interactions of individual cargo molecules with forming CCPs in living cells.
  • To investigate the binding and unbinding kinetics of cargo within CCPs and compare them to the overall endocytic timescale.

Main Methods:

  • Single-molecule total internal reflection fluorescence microscopy (TIRFm) was employed.
  • Automatic detection and tracking algorithms were used to monitor individual voltage-gated potassium channels.
  • Tens of thousands of cargo-CCP capturing events were analyzed.

Main Results:

  • Individual cargo molecules were observed associating with and dissociating from CCPs, with occasional internalization.
  • Cargo molecules frequently escaped CCPs before pit termination or vesicle formation, indicating short binding times.
  • Measured distributions of cargo capture and confinement times showed excellent agreement with a non-Markovian stochastic model.

Conclusions:

  • Cargo escape from CCPs is a significant factor contributing to CCP dynamic heterogeneity.
  • Short cargo binding times provide a mechanism for anomalous protein diffusion in the plasma membrane.
  • The findings challenge existing models of cargo-CCP interactions during endocytosis.