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Related Concept Videos

Vesicular Tubular Clusters01:45

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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.
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Proteins and neurotransmitters in secretory vesicles can be released from a cell upon vesicle docking, priming, and fusion with the plasma membrane. Vesicles are docked and primed in preparation for the quick exocytosis of their contents in response to a stimulus. The fusion process is mainly carried out by a SNAP Receptor or SNARE complex, consisting of synaptobrevin, syntaxin-1, and SNAP-25.
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SNAREs and Membrane Fusion01:43

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Once a transport vesicle has recognized its target organelle, the vesicular membrane needs to fuse with the target membrane to unload the cargo. Transmembrane proteins called SNAREs present on organelle membranes and their vesicles, mediate vesicle fusion.
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Cryo-electron Microscopy01:28

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Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...
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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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Intraluminal vesicles (ILVs) are small vesicles 50-80 nm in diameter formed during the maturation of early endosomes. A specialized endosome containing numerous ILVs is called a multivesicular body (MVB). ILVs contain internalized molecules such as antigens, nucleic acids, proteins, and metabolites. Some of these molecules are released from the MVBs inside exosomes and are transported to other cells. Other MVBs contain molecules that are retained in the ILVs and are later degraded within the...
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Freeze-Fracture Electron Microscopy for Extracellular Vesicle Analysis
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Extracellular vesicle fusion visualized by cryo-electron microscopy.

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  • 1Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot 7610001, Israel.

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Extracellular vesicles (EVs) deliver cargo via membrane fusion, similar to viruses. A new assay reveals EVs fuse dependently on pH and proteins, offering insights into their cellular delivery mechanisms.

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Area of Science:

  • Cell Biology
  • Biophysics
  • Virology

Background:

  • Extracellular vesicles (EVs) mediate intercellular communication by transferring bioactive molecules.
  • EV cargo delivery is hypothesized to involve protein-mediated, pH-dependent membrane fusion.
  • Current methods are insufficient to identify EV fusion proteins and elucidate their mechanisms.

Purpose of the Study:

  • To develop and validate an in vitro biophysical assay for investigating EV membrane fusion.
  • To identify key proteins and pH-dependent factors involved in EV fusion.
  • To visualize the membrane fusion process of EVs.

Main Methods:

  • Development and benchmarking of an in vitro biophysical assay for membrane fusion.
  • Standardization of the assay by comparing EV and viral fusion with liposomes.
  • Cryo-electron tomography for direct visualization of membrane fusion stages.

Main Results:

  • EVs and retroviruses exhibit pH- and protein-dependent fusion with late endosome-mimicking liposomes.
  • Cryo-electron tomography visualized the distinct stages of EV membrane fusion.
  • EVs maintain fusogenicity after acidification and reneutralization, unlike most retroviruses.

Conclusions:

  • The developed assay provides a robust method for studying EV membrane fusion.
  • EV fusion is a protein-mediated and pH-sensitive process.
  • EVs possess unique fusogenic properties compared to retroviruses, with implications for cargo delivery.