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Updated: Dec 22, 2025

Size Exclusion Chromatography to Analyze Bacterial Outer Membrane Vesicle Heterogeneity
Published on: March 31, 2021
Within-species variation in OMV cargo proteins: the Myxococcus xanthus OMV pan-proteome
Allison S Zwarycz1, Paul G Livingstone, David E Whitworth
1Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, Aberystwyth, Ceredigion SY23 4DD, UK. dew@aber.ac.uk.
Abstract:
Extracellular membrane vesicles are produced by all domains of life (bacteria, archaea and eukaryotes). Bacterial extracellular vesicles (outer membrane vesicles or OMVs) are produced by outer membrane blebbing, and contain proteins, nucleic acids, virulence factors, lipids and metabolites. OMV functions depend on their internal composition, therefore understanding the proteome of OMVs, and how it varies between organisms, is imperative. Here, we report a comparative proteomic profiling of OMVs from strains of Myxococcus xanthus, a predatory species of Gram-negative myxobacteria whose secretions include secondary metabolites and hydrolytic enzymes, thought to be involved in prey lysis. Ten strains were chosen for study, of which seven had genome sequences available. The remaining three strains were genome sequenced allowing definition of the core and accessory genes and genome-derived proteins found within the pan-genome and pan-proteome respectively. OMVs were isolated from each strain and proteins identified using mass spectrometry. The M. xanthus OMV pan-proteome was found to contain tens of 'core' and hundreds of 'accessory' proteins. Properties of the OMV pan-proteome were compared with those of the pan-proteome deduced from the M. xanthus pan-genome. On average, 80% of 'core' OMV proteins are encoded by genes of the core genome, yet the OMV proteomes of individual strains contain subsets of core genome-derived proteins which only partially overlap. In addition, the distribution of characteristics of vesicle proteins does not correlate with the genome-derived proteome characteristic distribution. We hypothesize that M. xanthus cells package a personalized subset of proteins whose availability is only partially dictated by the presence/absence of encoding genes within the genome.
Insights
Bacterial outer membrane vesicles (OMVs) contain diverse proteins. Myxococcus xanthus OMVs package a personalized protein subset, only partially influenced by genome-encoded genes, impacting their function.
Area of Science:
- Microbiology
- Proteomics
- Bacterial Physiology
Background:
- Extracellular membrane vesicles (EMVs) are produced by all life domains.
- Bacterial outer membrane vesicles (OMVs) contain proteins, nucleic acids, and metabolites, influencing their functions.
- Understanding OMV proteome variation is crucial for deciphering their roles.
Purpose of the Study:
- To perform comparative proteomic profiling of OMVs from Myxococcus xanthus strains.
- To define the pan-proteome of M. xanthus OMVs and compare it with the pan-genome.
- To investigate the relationship between genome-encoded proteins and OMV protein content.
Main Methods:
- Isolation and mass spectrometry-based proteomic analysis of OMVs from ten M. xanthus strains.
- Genome sequencing of three strains to define the pan-genome and pan-proteome.
- Comparative analysis of OMV proteomes, pan-proteome, and pan-genome.
Main Results:
- The M. xanthus OMV pan-proteome comprises 'core' and 'accessory' proteins.
- Approximately 80% of 'core' OMV proteins are encoded by the core genome.
- Individual OMV proteomes show partial overlap, and protein characteristics do not correlate with genome-derived proteome distributions.
Conclusions:
- M. xanthus cells appear to package a personalized subset of proteins into OMVs.
- The availability of OMV proteins is only partially dictated by the presence of encoding genes in the genome.
- This suggests a regulatory mechanism beyond simple gene presence/absence controls OMV composition.
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