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Related Concept Videos

Recycling Endosomes and Transcytosis00:58

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The recycling endosome, also known as the endosomal recycling compartment (ERC), is a part of the slow-recycling process of the endocytic pathway. Molecules internalized through receptor-mediated endocytosis are either degraded in the lysosomes or are recycled to the plasma membrane through the fast- or slow-recycling route.
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Essential proteins such as insulin or low-density lipoprotein (LDL) and micronutrients such as iron enter a eukaryotic cell through receptor-mediated endocytosis. Subsequently, the early endosomes fuse with the vesicles containing such receptor-ligand complexes and play a vital role in sorting the incoming ligands and receptors. While the ligands are either degraded inside the vesicle or released into the cytosol, their receptors are returned to the plasma membrane for further rounds of...
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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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Maturation of Endosomes01:28

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The early endosome containing internalized molecules matures through transformations in its location, morphology, intraluminal pH, and membrane protein composition. Together, these changes result in a more acidic late endosome that contains multiple intraluminal vesicles; therefore, the late endosome is also called a multivesicular body (MVB).
Changes in location
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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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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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Related Experiment Video

Updated: Jul 11, 2025

Author Spotlight: Development of a Large-Scale, Reproducible Production Method for Exosome Mimetics Using Magnetic Nanoparticles
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Design rules for efficient endosomal escape.

Madeline Zoltek1,2, Angel Vázquez2, Xizi Zhang2

  • 1Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720, USA.

Biorxiv : the Preprint Server for Biology
|November 14, 2023
PubMed
Summary
This summary is machine-generated.

ZF5.3 facilitates protein delivery by exploiting endosomal pathways. Cargo size and unfolding ease dictate efficiency, guiding the design of better therapeutic proteins.

Keywords:
Major: Biological SciencesMinor: Biochemistryfluorescence correlation spectroscopynanobodyprotein deliveryprotein foldingprotein therapeutics

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Area of Science:

  • Cell biology
  • Biochemistry
  • Protein engineering

Background:

  • Protein translocation across membranes is crucial for therapeutics but often inefficient.
  • Endosomal escape is a major bottleneck in protein delivery.
  • ZF5.3 is a mini-protein that aids protein translocation via endosomal pathways.

Approach:

  • Investigated the impact of cargo protein size and thermal stability on ZF5.3-mediated endosomal escape.
  • Utilized fluorescence correlation spectroscopy to quantify cytosolic protein concentration.
  • Analyzed the role of the homotypic fusion and protein sorting (HOPS) complex in the delivery pathway.

Key Points:

  • Protein delivery efficiency via ZF5.3 depends on both cargo size and its unfolding characteristics.
  • Low-melting temperature (Tm) proteins, including intrinsically disordered domains, utilize a HOPS-dependent high-efficiency escape route.
  • Small proteins are delivered moderately efficiently via the HOPS pathway, irrespective of their Tm.

Conclusions:

  • Identified a novel protein- and/or lipid-dependent endosomal escape pathway exploited by ZF5.3.
  • Findings provide design principles for enhancing therapeutic protein delivery by optimizing cargo properties.