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Proteomic analysis of bacterial extracellular vesicles enriched by ultracentrifugation and an ε-Poly- L-lysine-based method

ABSTRACT: Both Gram-negative and Gram-positive bacteria can release vesicle-like structures referred as bacterial extracellular vesicles (BEVs), which contain various bioactive compounds. BEVs play important roles in the microbial community interactions and host-microbe interactions. Markedly, BEVs can be delivered to host cell, thus modulating the development and function of the innate immune system. To clarify the compositions and biological functions of BEVs, these vesicles need to be collected with high purity and bioactivity. Here we propose an isolation strategy based on a broad-spectrum antimicrobial epsilon-poly-L–lysine (ε-PL) to precipitate BEVs at a relatively low centrifugal speed (10,000 × g). Compared to the standard ultracentrifugation strategy, our method can enrich BEVs from large volumes of media inexpensively and rapidly. The precipitated BEVs can be recovered by adjusting pH and ionic strength of the media, followed by ultrafiltration step to remove ε-PL and achieve buffer exchange. Protein compositions of the BEVs enriched by ultracentrifugation and ε-PL-based method are measured using LC-MS/MS. The resultant data indicate that protein compositions of the ε-PL-precipitated BEVs are comparable to those purified by ultracentrifugation.

INSTRUMENT(S): Q Exactive

ORGANISM(S): Escherichia Coli Staphylococcus Aureus

TISSUE(S): Cell Culture

SUBMITTER: Shujin Wei

LAB HEAD: Wanli Xing

PROVIDER: PXD034259 | Pride | 2022-08-03

REPOSITORIES: Pride

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