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PXD025393

PXD025393 is an original dataset announced via ProteomeXchange.

Dataset Summary
TitleIsolation method and characterization of outer membranes vesicles of Helicobacter pylori grown in a chemically defined medium
DescriptionOuter membrane vesicles (OMVs) are small vesicles constitutively shed by all Gram-negative bacterium, which have been proposed to play a role in Helicobacter pylori persistence and pathogenesis. The methods currently available for the isolation of H. pylori OMVs are diverse and time-consuming, raising the need for a protocol standardization, which was the main aim of this study. Here, we showed that the chemically defined F12 medium, supplemented with cholesterol, nutritionally supports bacterial growth and maintains H. pylori viability for at least 72 h. Additionally, we developed an abridged protocol for isolation of OMVs from these bacterial cultures, which comprises a low-speed centrifugation, supernatant filtration through a 0.45 µm pore, and two ultracentrifugations for OMVs’ recovery and washing. Using this approach, a good yield of highly pure bona fide OMVs was recovered from cultures of different H. pylori strains and in different periods of bacterial growth. Analysis of the proteome of OMVs isolated from H. pylori F12-cholesterol cultures validated the efficacy of our isolation protocol in attaining a higher sample purity with low representation of flagella proteins. The comparison with the proteomes of H. pylori OMVs available in the literature demonstrated that OMVs carry a selective protein cargo, suggesting their importance for H. pylori survival and pathogenesis. In conclusion, this work proposes a time- and cost-efficient protocol for the isolation of H. pylori OMVs from a chemically defined culture medium that is suitable for implementation in research and in the biopharmaceutical field.
HostingRepositoryPRIDE
AnnounceDate2021-05-31
AnnouncementXMLSubmission_2021-05-31_07:55:27.369.xml
DigitalObjectIdentifierhttps://dx.doi.org/10.6019/PXD025393
ReviewLevelPeer-reviewed dataset
DatasetOriginOriginal dataset
RepositorySupportSupported dataset by repository
PrimarySubmitterHugo Osorio
SpeciesList scientific name: Helicobacter pylori 26695; NCBI TaxID: 85962;
ModificationListiodoacetamide derivatized residue
InstrumentQ Exactive
Dataset History
RevisionDatetimeStatusChangeLog Entry
02021-04-14 04:27:35ID requested
12021-05-31 07:55:27announced
Publication List
Dataset with its publication pending
Keyword List
submitter keyword: Helicobacter pylori, bacterial outer membrane vesicles, defined medium, isolation protocol, proteomics, bacteria viability
Contact List
Ceu Figueiredo
contact affiliationi3S – Instituto de Investigação e Inovação em Saúde, Universidade do Porto, Porto, Portugal; Ipatimup – Instituto de Patologia e Imunologia Molecular da Universidade do Porto, Porto, Portugal;
contact emailcfigueiredo@ipatimup.pt
lab head
Hugo Osorio
contact affiliationi3S - Institute for Research and Innovation in Health, University of Porto
contact emailhosorio@i3s.up.pt
dataset submitter
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Dataset FTP location
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