Related Experiment Video

Updated: Aug 8, 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 27, 2011

Clint: a novel clathrin-binding ENTH-domain protein at the Golgi

Christoph Kalthoff1, Stephanie Groos, Rüdiger Kohl

  • 1Department of Cell Biology, Center of Anatomy, Hannover Medical School, Hannover, Germany.

Molecular Biology of the Cell
|November 14, 2002
PubMed

Insights

We identified Clint, a novel protein involved in clathrin-coated vesicle formation at the trans-Golgi network. Clint binds to clathrin and adaptors, localizing to Golgi and vesicles.

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Protein Interactions

Background:

  • Clathrin-mediated endocytosis is crucial for protein transport.
  • The trans-Golgi network (TGN) is a key sorting station for proteins.
  • Novel proteins involved in vesicle formation at the TGN are continuously being discovered.

Purpose of the Study:

  • To characterize a newly identified protein, Clint, and elucidate its role in clathrin-coated vesicle formation.
  • To investigate the binding partners and localization of Clint within epithelial cells.

Main Methods:

  • Protein characterization and binding assays (recombinant protein interactions).
  • Immunofluorescence microscopy to determine protein localization and colocalization.
  • Copurification studies with clathrin-coated vesicles.
  • Analysis of protein expression in rat kidney and small intestine.

Main Results:

  • Clint, a 68-kDa ENTH-domain protein, binds directly to clathrin, AP-1, beta2-adaptin, and alpha-adaptin.
  • Clint localizes to the Golgi region and peripheral vesicles, colocalizing with AP-1 and clathrin in the perinuclear area.
  • Clint's ENTH domain binds phosphoinositides, and it copurifies with clathrin-coated vesicles.
  • Clint is expressed in the apical region of kidney collecting duct and intestinal enterocytes.

Conclusions:

  • Clint is a novel clathrin-binding protein potentially involved in clathrin-coated vesicle formation at the TGN.
  • Clint may remain associated with vesicles longer than clathrin and adaptors.
  • Its unique methionine-rich domain warrants further investigation.

More Related Videos

Visualizing Clathrin-mediated Endocytosis of G Protein-coupled Receptors at Single-event Resolution via TIRF Microscopy12:40

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

Published on: October 20, 2014

Applications of pHluorin for Quantitative, Kinetic and High-throughput Analysis of Endocytosis in Budding Yeast10:02

Applications of pHluorin for Quantitative, Kinetic and High-throughput Analysis of Endocytosis in Budding Yeast

Published on: October 23, 2016

Related Concept Videos

COP Coated Vesicles00:59

COP Coated Vesicles

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 different...
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...
Coat Assembly and GTPases01:33

Coat Assembly and GTPases

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...
Vesicular Tubular Clusters01:45

Vesicular Tubular Clusters

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...
Transport Across the Golgi01:26

Transport Across the Golgi

While it is unclear how molecules move between adjacent Golgi cisternae, it is apparent that the molecules move from cis- cisterna, the entry face, to the trans- cisterna, the exit face. Experiments initially suggested vesicles that bud from one cisterna and fuse with the next cisterna to transport proteins between the cisternae. This vesicular transport model describes the Golgi apparatus as a relatively static structure with a unique enzyme composition in each cisterna. Molecules are...
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...