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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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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.
Coat assembly depends on the local availability of phosphatidylinositol phosphates or PIPs and GTP-binding proteins. Adaptor proteins, which link the coat proteins to the membrane, bind to these PIPs and play a crucial role in controlling...
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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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Vesicular Tubular Clusters01:45

Vesicular Tubular Clusters

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After budding out from the ER membrane, some COPII vesicles lose their coat and fuse with one another to form larger vesicles and interconnected tubules called vesicular tubular clusters or VTCs. These clusters constitute a compartment at the ER-Golgi interface known as ERGIC (Endoplasmic Reticulum Golgi Intermediate Compartment). The ERGIC is a mobile membrane-bound cargo transport system that sorts proteins secreted from ER and delivers them to the Golgi.
With the help of motor proteins such...
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Receptor-mediated Endocytosis01:39

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In vivo and in vitro Studies of Adaptor-clathrin Interaction
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In vivo and in vitro Studies of Adaptor-clathrin Interaction

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Multi-modal adaptor-clathrin contacts drive coated vesicle assembly.

Sarah M Smith1, Gabrielle Larocque2, Katherine M Wood1

  • 1School of Life Sciences, University of Warwick, Coventry, UK.

The EMBO Journal
|September 6, 2021
PubMed
Summary

The AP2 complex binds clathrin triskelia in multiple ways during endocytosis. This multi-modal binding, involving specific sites on the AP2 appendage and clathrin, is crucial for forming coated pits.

Keywords:
clathrincryo-electron microscopyendocytosismembrane traffic

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Area of Science:

  • Cell biology
  • Structural biology
  • Biochemistry

Background:

  • Clathrin-coated pits mediate endocytosis, a vital cellular process.
  • The AP2 complex initiates clathrin recruitment, but its role in subsequent assembly is unclear.
  • Structural and functional details of AP2-clathrin interactions are incomplete.

Purpose of the Study:

  • To investigate the structural basis of AP2-clathrin interactions during coated-pit assembly.
  • To elucidate the functional significance of these interactions for endocytosis.

Main Methods:

  • Cryo-electron microscopy (cryo-EM) to determine the structure of clathrin cages assembled with AP2 beta2 hinge-appendage (beta2HA).
  • Biochemical analysis of beta2HA-clathrin interactions.
  • Functional assays to assess the role of identified interaction sites in endocytosis.

Main Results:

  • The cryo-EM structure revealed that the beta2-appendage binds clathrin cages in at least two distinct positions.
  • One binding mode involves the beta2-appendage cross-linking terminal domains of adjacent clathrin triskelia.
  • Functional studies identified two essential clathrin interaction sites: a clathrin-box motif on the hinge and a "sandwich site" on the appendage.

Conclusions:

  • AP2-clathrin interactions are multi-modal, with the beta2-appendage binding to multiple triskelia.
  • This multi-site binding is a fundamental property driving clathrin cage assembly.
  • Both hinge and appendage interaction sites on AP2 are critical for efficient endocytosis.