PXD025393

PXD025393 is an original dataset announced via ProteomeXchange.

Title	Isolation method and characterization of outer membranes vesicles of Helicobacter pylori grown in a chemically defined medium
Description	Outer 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.
HostingRepository	PRIDE
AnnounceDate	2021-05-31
AnnouncementXML	Submission_2021-05-31_07:55:27.369.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD025393
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	Hugo Osorio
SpeciesList	scientific name: Helicobacter pylori 26695; NCBI TaxID: 85962;
ModificationList	iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2021-04-14 04:27:35	ID requested
⏵ 1	2021-05-31 07:55:27	announced

Dataset with its publication pending

submitter keyword: Helicobacter pylori, bacterial outer membrane vesicles, defined medium, isolation protocol, proteomics, bacteria viability

Ceu Figueiredo
contact affiliation	i3S – 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 email	cfigueiredo@ipatimup.pt
lab head
Hugo Osorio
contact affiliation	i3S - Institute for Research and Innovation in Health, University of Porto
contact email	hosorio@i3s.up.pt
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD025393
     1. Label: PRIDE project
     2. Name: Isolation method and characterization of outer membranes vesicles of Helicobacter pylori grown in a chemically defined medium