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Related Concept Videos

Gram-negative Bacterial Protein Secretion Systems01:17

Gram-negative Bacterial Protein Secretion Systems

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Gram-negative bacteria utilize sophisticated protein secretion systems to transport proteins across their double-membrane envelope into the extracellular environment or host cells. Based on their mechanism of action, these systems are classified into one-step and two-step pathways.One-Step Secretion Systems (Types I, III, IV, and VI)One-step secretion systems bypass the periplasm entirely, forming a continuous channel that spans both the inner and outer membranes:Type I Secretion System (T1SS):...
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Bacterial protein secretion involves translocation systems to ensure proteins reach their designated locations, including the plasma membrane, periplasm, outer membrane, or the external environment. These translocation systems are vital for bacterial physiology, supporting processes like membrane assembly, enzymatic activity in the periplasm, and interactions with the external environment. The division of labor between Sec and Tat pathways ensures efficiency in handling proteins with diverse...
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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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Secretory vesicles, also known as dense core vesicles (DCVs), are membrane-bound vesicles that transport secretory proteins, such as hormones or neurotransmitters. Regulated secretory vesicles transport proteins from the trans-Golgi network to the exterior of the cell. Proteins present in regulated secretory vesicles are required to be rapidly exocytosed in large amounts upon a specific stimulus.
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Eukaryotic cells have different membrane-bound organelles with distinct protein requirements. The process by which proteins are targeted to a specific organelle is called protein sorting.
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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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Updated: Jul 29, 2025

Size Exclusion Chromatography to Analyze Bacterial Outer Membrane Vesicle Heterogeneity
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Identifying size-dependent toxin sorting in bacterial outer membrane vesicles.

Aarshi N Singh1, Justin B Nice2, Angela C Brown2

  • 1Department of Chemistry, Lehigh University, Bethlehem, PA, U.S.A.

Biorxiv : the Preprint Server for Biology
|May 19, 2023
PubMed
Summary
This summary is machine-generated.

Gram-negative bacteria release outer membrane vesicles (OMVs) with varying toxin levels. Larger OMVs from Aggregatibacter actinomycetemcomitans are more likely to contain leukotoxin (LtxA).

Keywords:
Single particle sizingfluorescent microscopyouter membrane vesiclestoxins

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Area of Science:

  • Microbiology
  • Nanotechnology
  • Biochemistry

Background:

  • Gram-negative bacteria utilize outer membrane vesicles (OMVs) for intercellular communication and virulence.
  • Heterogeneity in OMV size and toxin content complicates ensemble-based analyses.
  • Understanding OMV heterogeneity is crucial for deciphering bacterial pathogenesis.

Approach:

  • Employed single OMV fluorescence imaging to resolve nanoscale surface heterogeneity.
  • Quantified size-dependent leukotoxin (LtxA) sorting in OMVs from Aggregatibacter actinomycetemcomitans (A.a.).
  • Developed a non-invasive method to assess OMV composition without prior separation.

Key Points:

  • Aggregatibacter actinomycetemcomitans (A.a.) produces OMVs with a bimodal size distribution.
  • Leukotoxin (LtxA) presence significantly correlates with OMV size.
  • Small OMVs (<100 nm) show 0-30% LtxA positivity, while large OMVs (>200 nm) exhibit 70-100% LtxA positivity.

Conclusions:

  • Single OMV imaging reveals nanoscale OMV surface heterogeneity.
  • Demonstrated size-based sorting of leukotoxin (LtxA) in bacterial OMVs.
  • This technique bypasses the need for OMV fraction separation for heterogeneity analysis.