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- Title
Loss of Lipooligosaccharide Synthesis in Acinetobacter baumannii Produces Changes in Outer Membrane Vesicle Protein Content.
- Authors
Cano-Castaño, Beatriz; Corral-Lugo, Andrés; Gato, Eva; Terrón, María C.; Martín-Galiano, Antonio J.; Sotillo, Javier; Pérez, Astrid; McConnell, Michael J.
- Abstract
Outer membrane vesicles (OMVs) are nanostructures derived from the outer membrane of Gram-negative bacteria. We previously demonstrated that vaccination with endotoxin-free OMVs isolated from an Acinetobacter baumannii strain lacking lipooligosaccharide (LOS) biosynthesis, due to a mutation in lpxD, provides full protection in a murine sepsis model. The present study characterizes the protein content of highly-purified OMVs isolated from LOS-replete and LOS-deficient strains. Four purification methods were evaluated to obtain highly purified OMV preparations: ultracentrifugation, size exclusion chromatography (SEC), ultracentrifugation followed by SEC, and Optiprep™. OMVs from each method were characterized using nanoparticle tracking analysis and electron microscopy. OMVs from LOS-deficient and LOS-replete strains purified using the Optiprep™ method were subjected to LC-MS/MS analysis to determine protein content. Significant differences in protein composition between OMVs from LOS-deficient and LOS-replete strains were found. Computational analyses using Bepipred 3.0 and SEMA 2.0 indicated that the lack of LOS led to the overexpression of immunogenic proteins found in LOS-containing OMVs and the presence of immune-stimulating proteins absent in LOS-replete OMVs. These findings have important implications for developing OMV-based vaccines against A. baumannii, using both LOS-containing and LOS-free OMVs preparations.
- Subjects
EXTRACELLULAR vesicles; GEL permeation chromatography; ACINETOBACTER baumannii; PROTEIN overexpression; GRAM-negative bacteria
- Publication
International Journal of Molecular Sciences, 2024, Vol 25, Issue 17, p9272
- ISSN
1661-6596
- Publication type
Article
- DOI
10.3390/ijms25179272