Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Concept Videos

The Movement of Organelles and Vesicles01:43

The Movement of Organelles and Vesicles

In eukaryotic cells,  cytoskeletal filaments such as actin, microtubules, and intermediate filaments form a mesh-like cytoskeletal network. These filaments serve as tracks for transporting cellular cargo. Specialized motor proteins use the chemical energy stored in adenosine triphosphate (ATP) for this transport. During interphase, microtubules are polarized, with the plus-end towards the cell periphery and the minus-end towards the cell center. Two microtubule-associated motor proteins,...
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...

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

A discrete choice experiment investigating HIV testing preferences in South Africa.

Journal of medical economics·2022
Same author

Gender (in) differences in prevalence and incidence of traumatic experiences among orphaned and separated children living in five low- and middle-income countries.

Global mental health (Cambridge, England)·2015
Same author

Perceived acceptability of home-based couples voluntary HIV counseling and testing in Northern Tanzania.

AIDS care·2011
Same author

Low sensitivity of T-cell based detection of tuberculosis among HIV co-infected Tanzanian in-patients.

East African medical journal·2009
Same author

Morbidity and mortality among a cohort of HIV-infected adults in a programme for community home-based care, in the Kilimanjaro Region of Tanzania (2003-2005).

Annals of tropical medicine and parasitology·2009
Same author

Trauma, anxiety and reported health among HIV-positive persons in Tanzania and the US Deep South.

AIDS care·2008
Same journal

A viral ORFeome library for systems-level genetic dissection of host-pathogen interactions.

Cell·2026
Same journal

Co-option of lysosomal machinery shapes the evolution of the intracellular photosymbiosis supporting coral reefs.

Cell·2026
Same journal

LEF1 and niche factors determine T cell stemness across chronic diseases.

Cell·2026
Same journal

Recurrent patterns of TOP1-mediated neuronal genomic damage shared by major neurodegenerative disorders.

Cell·2026
Same journal

Four-dimensional molecular mapping from a spatial snapshot reveals the dynamics of hair follicle organogenesis.

Cell·2026
Same journal

Whole-cell particle-based digital twin simulations from 4D lattice light-sheet microscopy data.

Cell·2026
See all related articles

Related Experiment Video

Updated: Jun 16, 2026

Visualizing Intracellular SNARE Trafficking by Fluorescence Lifetime Imaging Microscopy
08:55

Visualizing Intracellular SNARE Trafficking by Fluorescence Lifetime Imaging Microscopy

Published on: December 29, 2017

Stepwise assembly of functionally active transport vesicles

J Ostermann1, L Orci, K Tani

  • 1Cellular Biochemistry and Biophysics Program Memorial Sloan Kettering Cancer Center, New York, New York 10021.

Cell
|December 3, 1993
PubMed
Summary
This summary is machine-generated.

Researchers identified the essential components and steps for forming COP-coated vesicles, crucial for protein transport. This study clarifies the in vitro generation of these vesicles, aiding in understanding intracellular protein trafficking.

More Related Videos

Directed Assembly of Elastin-like Proteins into defined Supramolecular Structures and Cargo Encapsulation In Vitro
10:01

Directed Assembly of Elastin-like Proteins into defined Supramolecular Structures and Cargo Encapsulation In Vitro

Published on: April 8, 2020

Construction of Out-of-Equilibrium Metabolic Networks in Nano- and Micrometer-Sized Vesicles
10:56

Construction of Out-of-Equilibrium Metabolic Networks in Nano- and Micrometer-Sized Vesicles

Published on: April 12, 2024

Related Experiment Videos

Last Updated: Jun 16, 2026

Visualizing Intracellular SNARE Trafficking by Fluorescence Lifetime Imaging Microscopy
08:55

Visualizing Intracellular SNARE Trafficking by Fluorescence Lifetime Imaging Microscopy

Published on: December 29, 2017

Directed Assembly of Elastin-like Proteins into defined Supramolecular Structures and Cargo Encapsulation In Vitro
10:01

Directed Assembly of Elastin-like Proteins into defined Supramolecular Structures and Cargo Encapsulation In Vitro

Published on: April 8, 2020

Construction of Out-of-Equilibrium Metabolic Networks in Nano- and Micrometer-Sized Vesicles
10:56

Construction of Out-of-Equilibrium Metabolic Networks in Nano- and Micrometer-Sized Vesicles

Published on: April 12, 2024

Area of Science:

  • Cell Biology
  • Molecular Biology
  • Biochemistry

Background:

  • COP-coated vesicles mediate protein transport from the endoplasmic reticulum through the Golgi apparatus.
  • Their formation involves cytosolic factors like ADP-ribosylation factor (ARF) and coatomer proteins, along with GTP and fatty acyl-coenzyme A (CoA).

Purpose of the Study:

  • To elucidate the minimal cytosolic requirements for COP-coated vesicle budding from Golgi cisternae.
  • To characterize the stepwise assembly and in vitro generation of functional COP-coated vesicles.

Main Methods:

  • In vitro reconstitution assays using Golgi membranes and purified cytosolic components.
  • Stepwise addition of ADP-ribosylation factor (ARF), coatomer, GTP, and palmitoyl-CoA.
  • Isolation and functional assessment of generated COP-coated vesicles.

Main Results:

  • Identified ADP-ribosylation factor (ARF), coatomer, GTP, and fatty acyl-coenzyme A (CoA) as essential for vesicle budding.
  • Demonstrated that coatomer, ARF, and GTP are required for coated bud assembly.
  • Showed that palmitoyl-CoA addition triggers membrane fission, releasing functional coated vesicles.
  • Established that COP-coated vesicles can be generated stepwise in vitro and isolated in an active state.

Conclusions:

  • The minimal set of cytosolic components and principal steps for COP-coated vesicle formation have been identified.
  • This work provides a foundation for further investigation into the mechanisms of intracellular protein transport and vesicle trafficking.