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Identifying size-dependent toxin sorting in bacterial outer membrane vesicles

Gram-negative bacteria produce outer membrane vesicles (OMVs) that play a critical role in cell-cell communication and virulence. Despite being isolated from a single population of bacteria, OMVs can exhibit heterogeneous size and toxin content, which can be obscured by assays that measure ensemble...

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Autores principales: Singh, Aarshi N., Nice, Justin B, Brown, Angela C., Wittenberg, Nathan J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10187208/
https://www.ncbi.nlm.nih.gov/pubmed/37205353
http://dx.doi.org/10.1101/2023.05.03.539273
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author Singh, Aarshi N.
Nice, Justin B
Brown, Angela C.
Wittenberg, Nathan J.
author_facet Singh, Aarshi N.
Nice, Justin B
Brown, Angela C.
Wittenberg, Nathan J.
author_sort Singh, Aarshi N.
collection PubMed
description Gram-negative bacteria produce outer membrane vesicles (OMVs) that play a critical role in cell-cell communication and virulence. Despite being isolated from a single population of bacteria, OMVs can exhibit heterogeneous size and toxin content, which can be obscured by assays that measure ensemble properties. To address this issue, we utilize fluorescence imaging of individual OMVs to reveal size-dependent toxin sorting. Our results showed that the oral bacterium Aggregatibacter actinomycetemcomitans (A.a.) produces OMVs with a bimodal size distribution, where larger OMVs were much more likely to possess leukotoxin (LtxA). Among the smallest OMVs (< 100 nm diameter), the fraction that are toxin positive ranges from 0–30%, while the largest OMVs (> 200 nm diameter) are between 70–100% toxin positive. Our single OMV imaging method provides a non-invasive way to observe OMV surface heterogeneity at the nanoscale level and determine size-based heterogeneities without the need for OMV fraction separation.
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spelling pubmed-101872082023-05-17 Identifying size-dependent toxin sorting in bacterial outer membrane vesicles Singh, Aarshi N. Nice, Justin B Brown, Angela C. Wittenberg, Nathan J. bioRxiv Article Gram-negative bacteria produce outer membrane vesicles (OMVs) that play a critical role in cell-cell communication and virulence. Despite being isolated from a single population of bacteria, OMVs can exhibit heterogeneous size and toxin content, which can be obscured by assays that measure ensemble properties. To address this issue, we utilize fluorescence imaging of individual OMVs to reveal size-dependent toxin sorting. Our results showed that the oral bacterium Aggregatibacter actinomycetemcomitans (A.a.) produces OMVs with a bimodal size distribution, where larger OMVs were much more likely to possess leukotoxin (LtxA). Among the smallest OMVs (< 100 nm diameter), the fraction that are toxin positive ranges from 0–30%, while the largest OMVs (> 200 nm diameter) are between 70–100% toxin positive. Our single OMV imaging method provides a non-invasive way to observe OMV surface heterogeneity at the nanoscale level and determine size-based heterogeneities without the need for OMV fraction separation. Cold Spring Harbor Laboratory 2023-05-09 /pmc/articles/PMC10187208/ /pubmed/37205353 http://dx.doi.org/10.1101/2023.05.03.539273 Text en https://creativecommons.org/licenses/by-nc-nd/4.0/This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which allows reusers to copy and distribute the material in any medium or format in unadapted form only, for noncommercial purposes only, and only so long as attribution is given to the creator.
spellingShingle Article
Singh, Aarshi N.
Nice, Justin B
Brown, Angela C.
Wittenberg, Nathan J.
Identifying size-dependent toxin sorting in bacterial outer membrane vesicles
title Identifying size-dependent toxin sorting in bacterial outer membrane vesicles
title_full Identifying size-dependent toxin sorting in bacterial outer membrane vesicles
title_fullStr Identifying size-dependent toxin sorting in bacterial outer membrane vesicles
title_full_unstemmed Identifying size-dependent toxin sorting in bacterial outer membrane vesicles
title_short Identifying size-dependent toxin sorting in bacterial outer membrane vesicles
title_sort identifying size-dependent toxin sorting in bacterial outer membrane vesicles
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10187208/
https://www.ncbi.nlm.nih.gov/pubmed/37205353
http://dx.doi.org/10.1101/2023.05.03.539273
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