Related Experiment Video
Updated: May 1, 2026
07:26
Size Exclusion Chromatography to Analyze Bacterial Outer Membrane Vesicle Heterogeneity
Published on: March 31, 2021
4.7K
Proteomic analysis of Vibrio cholerae outer membrane vesicles
Emrah Altindis1, Yang Fu, John J Mekalanos
1Department of Microbiology and Immunobiology, Harvard Medical School, Boston, MA 02115.
Summary
Outer membrane vesicles (OMVs) from Vibrio cholerae contain proteins crucial for bacterial survival and pathogenesis. The DegP protease significantly influences OMV content and bacterial intestinal colonization.
Area of Science:
- Microbiology
- Bacterial Pathogenesis
- Proteomics
Background:
- Outer membrane vesicles (OMVs) are key to Gram-negative bacteria's survival and pathogenesis.
- OMVs offer a method to study cell-envelope proteins without cell lysis.
- They are also explored as potential immunogenic platforms.
Purpose of the Study:
- To identify proteins within Vibrio cholerae OMVs under ToxT activation.
- To assess the virulence contribution of OMV-associated proteins.
- To investigate the role of DegP protease in OMV composition and function.
Main Methods:
- In-solution trypsin digestion coupled with mass spectrometry for OMV protein identification.
- ToxT expression analysis, in vivo RNA-seq, and Tn-seq for virulence assessment.
- Comparative proteomics of wild-type and degP mutant strains.
Main Results:
- Identified 90 proteins in V. cholerae OMVs under ToxT activation.
- Thirteen OMV proteins are essential for cell growth, suggesting potential drug targets.
- Twelve nonessential OMV proteins, including DegP, are vital for intestinal colonization.
- DegP influences the incorporation of nine proteins into OMVs, impacting biofilm formation and type II secretion.
Conclusions:
- DegP protease is critical for determining OMV protein content in V. cholerae.
- OMV composition, influenced by DegP, affects bacterial intestinal colonization.
- DegP impacts the type II secretion system and biofilm matrix formation.
Related Concept Videos
01:26Bacterial Translocation and Protein Secretion
1.1K
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...
1.1K
01:25Cholera
149
Cholera is an acute gastrointestinal disease caused by the Gram-negative bacterium Vibrio cholerae. It is transmitted primarily via the fecal-oral route through the ingestion of contaminated water or food.Vibrio cholerae is a motile, Gram-negative bacterium of the family Vibrionaceae, primarily associated with waterborne outbreaks in areas with inadequate sanitation. Although over 200 serogroups of V. cholerae exist, only O1 and O139 are responsible for epidemic cholera. The O1 serogroup,...
149