Department of Microbiology, Anatomy, Physiology and Pharmacology, La Trobe University, Bundoora, Australia.
Research Centre for Extracellular Vesicles, La Trobe University, Bundoora, Australia.
Proteomics. 2023 May;23(10):e2200464. doi: 10.1002/pmic.202200464. Epub 2023 Mar 17.
Gram-negative bacteria produce outer membrane vesicles (OMVs) and contain bacterial cargo including nucleic acids and proteins. The proteome of OMVs can be altered by various factors including bacterial growth stage, growth conditions, and environmental factors. However, it is currently unknown if the mechanism of OMV biogenesis can determine their proteome. In this study, we examined whether the mechanisms of OMV biogenesis influenced the production and protein composition of Pseudomonas aeruginosa OMVs. OMVs were isolated from three P. aeruginosa strains that produced OMVs either by budding alone, by explosive cell lysis, or by both budding and explosive cell lysis. We identified that the mechanism of OMV biogenesis dictated OMV quantity. Furthermore, a global proteomic analysis comparing the proteome of OMVs to their parent bacteria showed significant differences in the identification of proteins in bacteria and OMVs. Finally, we determined that the mechanism of OMV biogenesis influenced the protein composition of OMVs, as OMVs released by distinct mechanisms of biogenesis differed significantly from one another in their proteome and functional enrichment analysis. Overall, our findings reveal that the mechanism of OMV biogenesis is a main factor that determines the OMV proteome which may affect their subsequent biological functions.
革兰氏阴性菌会产生外膜囊泡（OMV），其中包含细菌货物，包括核酸和蛋白质。OMV 的蛋白质组可以通过多种因素改变，包括细菌生长阶段、生长条件和环境因素。然而，目前尚不清楚 OMV 生物发生的机制是否可以决定其蛋白质组。在这项研究中，我们研究了 OMV 生物发生的机制是否会影响铜绿假单胞菌 OMV 的产生和蛋白质组成。从通过出芽单独产生 OMV、通过爆炸性细胞裂解产生 OMV 或通过出芽和爆炸性细胞裂解两者产生 OMV 的三种铜绿假单胞菌菌株中分离出 OMV。我们发现 OMV 生物发生的机制决定了 OMV 的数量。此外，将 OMV 的蛋白质组与它们的亲代细菌进行比较的全蛋白质组分析表明，在细菌和 OMV 中鉴定蛋白质存在显着差异。最后，我们确定 OMV 生物发生的机制影响 OMV 的蛋白质组成，因为通过不同生物发生机制释放的 OMV 在蛋白质组和功能富集分析方面存在显着差异。总的来说，我们的研究结果表明，OMV 生物发生的机制是决定 OMV 蛋白质组的主要因素，这可能会影响它们随后的生物学功能。
