Dataset Information

Analysis of the Escherichia coli membrane vesicle proteome identifies markers of purity and culture conditions

ABSTRACT: Bacteria release nano-sized membrane vesicles (MVs) into the extracellular milieu. Bacterial MVs contain molecular cargo originating from the parent bacterium and have important roles in bacterial survival and pathogenesis. Using 8-plex iTRAQ approaches, we profiled the MV proteome of two Escherichia coli strains - uropathogenic E. coli (UPEC) 536 and probiotic Nissle 1917. For these strains, we compared the difference in the MV proteome between a crude input MV prepared by ultracentrifugation alone with that from MVs that were further purified by either density gradient centrifugation (DGC) or size exclusion chromatography (SEC); and also compared MVs from bacterial cultures that were grown in iron-restricted (R) and ironsupplemented (RF) conditions. We found that overall, outer membrane components were highly enriched, and bacterial inner membrane components were significantly depleted in both UPEC and Nissle MVs, in keeping with an outer membrane origin. In addition, we found enrichment of ribosome-related Gene Ontology terms in UPEC MV, and proteins involved in glycolytic process and ligase activity in Nissle MV. We have identified that three proteins (RbsB of UPEC in R; YoeA of UPEC in RF; BamA of Nissle in R) were consistently enriched in the DGC- and SEC-purified MV samples in comparison to their crude input MV, whereas conversely the 60 kDa chaperonin GroEL was enriched in the crude input MVs for both UPEC and Nissle in R condition. Such proteins may have a potential utility as technical markers for assessing the purity from contaminating non-vesicular protein after MV purification. Several proteins were changed in their abundance depending on the iron availability in the media.

INSTRUMENT(S): TripleTOF 6600

ORGANISM(S): Escherichia Coli

TISSUE(S): Prokaryotic Cell

DISEASE(S): Wounds And Injuries

SUBMITTER: Leo Payne

LAB HEAD: Simon Swift

PROVIDER: PXD011345 | Pride | 2019-07-15

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	170314_JH_Bact_memb_iTRAQ_2.group	Other
	170314_JH_Bact_memb_iTRAQ_2.wiff	Wiff
	170314_JH_Bact_memb_iTRAQ_2.wiff.scan	Wiff
	170328_Bact_vessicle_iTRAQ_1.group	Other
	170328_Bact_vessicle_iTRAQ_1.wiff	Wiff

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Publications

Analysis of the <i>Escherichia coli</i> extracellular vesicle proteome identifies markers of purity and culture conditions.

Hong Jiwon J Dauros-Singorenko Priscila P Whitcombe Alana A Payne Leo L Blenkiron Cherie C Phillips Anthony A Swift Simon S

Journal of extracellular vesicles 20190624 1

Bacteria release nano-sized extracellular vesicles (EVs) into the extracellular milieu. Bacterial EVs contain molecular cargo originating from the parent bacterium and have important roles in bacterial survival and pathogenesis. Using 8-plex iTRAQ approaches, we profiled the EV proteome of two <i>Escherichia coli</i> strains, uropathogenic (UPEC) 536 and probiotic Nissle 1917. For these strains, we compared the proteome of crude input EVs prepared by ultracentrifugation alone with EVs purified b ...[more]

PMID: 31275533

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