PXD022786 is an
original dataset announced via ProteomeXchange.
Dataset Summary
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 |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2020-11-27 06:10:14 | ID requested | |
⏵ 1 | 2021-09-09 14:17:19 | announced | |
Publication List
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] |
Keyword List
submitter keyword: Escherichia coli, OMV, TolA, LC-MSMS |
Contact List
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 | |
Full Dataset Link List
Dataset FTP location
NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2021/09/PXD022786 |
PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD022786
- Label: PRIDE project
- Name: Proteomic analysis of outer membrane vesicle isolation methods with an Escherichia coli tolA mutant using LC MSMS