Dataset Information

Proteome of mass-produced Escherichia coli outer membrane vesicles

ABSTRACT: Large-scale production of bacterial extracellular vesicles is a hurdle for their development as novel cancer immunotherapeutic agents. Here, we developed manufacturing processes for mass production of Escherichia coli EVs known as outer membrane vesicles (OMVs). We present a comprehensive proteome of mass-produced E. coli OMVs.

INSTRUMENT(S): Orbitrap Eclipse

ORGANISM(S): Escherichia Coli Bacteria

TISSUE(S): Cell Culture

DISEASE(S): Disease Free

SUBMITTER: Yong Song Gho

LAB HEAD: Yong Song Gho

PROVIDER: PXD043119 | Pride | 2023-10-24

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	EcoliOMV_01.mzML	Mzml
	EcoliOMV_01_interact.pep.xml	Pepxml
	EcoliOMV_01_interact.prot.xml	Xml
	EcoliOMV_02.mzML	Mzml
	EcoliOMV_02_interact.pep.xml	Pepxml

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Publications

Mass-produced gram-negative bacterial outer membrane vesicles activate cancer antigen-specific stem-like CD8<sup>+</sup> T cells which enables an effective combination immunotherapy with anti-PD-1.

Won Solchan S Lee Changjin C Bae Seoyoon S Lee Jaemin J Choi Dongsic D Kim Min-Gang MG Song Sunghyun S Lee Jaewook J Kim Eunhye E Shin HaYoung H Basukala Anita A Lee Tae Ryong TR Lee Dong-Sup DS Gho Yong Song YS

Journal of extracellular vesicles 20230801 8

Despite the capability of extracellular vesicles (EVs) derived from Gram-negative and Gram-positive bacteria to induce potent anti-tumour responses, large-scale production of bacterial EVs remains as a hurdle for their development as novel cancer immunotherapeutic agents. Here, we developed manufacturing processes for mass production of Escherichia coli EVs, namely, outer membrane vesicles (OMVs). By combining metal precipitation and size-exclusion chromatography, we isolated 357 mg in total pro ...[more]

PMID: 37563797

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Project description:Tenacibaculum maritimum, the etiological agent of tenacibaculosis in marine fish, constitutively secretes extracellular products (ECPs), which play a main role in virulence. However, their protein content has not been yet comprehensively studied. In this study a collection of 64 T. maritimum strains belonging to the four serotypes described so far (O1 to O4) was used to analyse the prevalence of extracellular proteolytic and lipolytic activities related to virulence. Results showed the existence of a large heterogeneity of the enzymatic capacity among serotype O4 isolates. Most notably, the ECPs of T. maritimum SP9.1 belonging to serotype O4 contain a large amount of outer membrane vesicles (OMVs), which were characterized by electron microscopy and purified by successive steps of filtration and centrifugation. Total protein content of ECPs, and the proteins associated to OMVs and soluble fraction of the ECPs (S-ECPs) were identified by nLC-TIMS-QTOF. A total of 641 proteins were identified in ECPs including some virulence related factors, which were mainly found in one of the fractions, either in OMVs or S-ECPs. Outer membrane proteins such as TonB-dependent transporters and the T9SS-related proteins PorP, PorT and SprA appeared to be mainly associated with OMVs. By contrast, putative virulence factors such as sialidase SiaA, chondroitinase CslA, sphingomyelinase Sph, ceramidase Cer and collagenase Col were found only in the S-ECPs. These findings demonstrate that T. maritimum release, through surface blebbing, outer membrane vesicles that are enriched specifically in TonB-dependent siderophore transporters and proteins of the type IX secretion system. Interestingly, our results also showed that OMVs could play a role in virulence by promoting surface adhesion, biofilm formation, and maximizing cytotoxic effects of the ECPs. The T. maritimum secretome analysis provides insights into the extracellular products function and can constitute the basis for future studies aimed to elucidate the role of OMVs in the pathogenesis of fish tenacibaculosis.