PXD022786

PXD022786 is an original dataset announced via ProteomeXchange.

Title	Proteomic analysis of outer membrane vesicle isolation methods with an Escherichia coli tolA mutant using LC MSMS
Description	Outer membrane vesicles (OMVs) produced by Gram-negative bacteria are mediators of cell survival and pathogenesis by facilitating nutrient transport, virulence factor dissemination, and resistance to antimicrobials. Studies of OMV properties often focus on hypervesiculating Escherichia coli mutants that have increased OMV production when compared to their corresponding wild-type (WT) strains. Currently, two conventional techniques, ultracentrifugation (UC) and ultradiafiltration (UF), are used interchangeably to isolate OMVs, however, there is concern that each technique may inadvertently alter the properties of isolated OMVs during study. To address this concern, we compared two OMV isolation methods, UC and UF, with respect to final OMV quantities, size distributions, and morphologies using a hypervesiculating Escherichia coli K-12 ∆tolA mutant. Cryo-transmission electron microscopy (cryo-TEM) visualization of isolated OMVs revealed distinct morphological differences between wild-type and ∆tolA OMVs, where ∆tolA OMVs isolated by either UC or UF method possessed a greater proportion of OMVs with two or more membranes (39.6-42.4%). Proteomic OMV analysis of WT and ∆tolA OMVs confirmed that ∆tolA enhances inner plasma membrane carryover in multi-lamellar OMVs. This study demonstrates that UC and UF are useful techniques for OMV isolation, where UF may be preferable due to faster isolation, higher OMV yields and enrichment of smaller sized vesicles.
HostingRepository	PRIDE
AnnounceDate	2021-09-09
AnnouncementXML	Submission_2021-09-09_14:17:17.642.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD022786
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Denice Bay
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: 562;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	LTQ Orbitrap Velos

Revision	Datetime	Status	ChangeLog Entry
0	2020-11-27 06:10:14	ID requested
⏵ 1	2021-09-09 14:17:19	announced

Reimer SL, Beniac DR, Hiebert SL, Booth TF, Chong PM, Westmacott GR, Zhanel GG, Bay DC, Mutant Reveals a Hypervesiculating Phenotype With Outer-Inner Membrane Vesicle Content. Front Microbiol, 12():628801(2021) [pubmed]

submitter keyword: Escherichia coli, OMV, TolA, LC-MSMS

Denice C Bay
contact affiliation	Department of Medical Microbiology and Infectious Diseases University of Manitoba 745 Bannatyne Avenue Winnipeg, MB, Canada, R3E 0J9
contact email	Denice.Bay@umanitoba.ca
lab head
Denice Bay
contact affiliation	The University of Manitoba, Department of Medical Microbiology and Infectious Diseases
contact email	Denice.Bay@umanitoba.ca
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD022786
     1. Label: PRIDE project
     2. Name: Proteomic analysis of outer membrane vesicle isolation methods with an Escherichia coli tolA mutant using LC MSMS