PXD075198-2

PXD075198 is an original dataset announced via ProteomeXchange.

Title	Lyophilization prior to homogenisation and extraction increases membrane protein detection in Gram-negative bacterial proteomic analyses
Description	Multi-drug resistant Gram-negative bacteria (GNB) are major contributors to the anti-microbial resistance (AMR) burden in humans and animals. AMR mechanisms are primarily mediated by proteoforms and, therefore, proteomic analyses of GNB offers a significant advantage in understanding the mechanisms of AMR. A large portion of these mechanisms are mediated by membrane proteins, however, they are often difficult to extract due to their hydrophobic nature and complex interactions with other components of the cell membrane. To extract the greatest number of proteoforms, an efficient ho-mogenisation protocol is required to effectively disrupt the rigid cell wall and membrane. Using Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumannii and Pseudomonas aeruginosa we systematically compared the extraction efficiency of bead-beating with flash frozen and lyophilized cell pellets. We demonstrate that lyophilization prior to ho-mogenisation by bead-beating increases the detection of hydrophobic, membrane pro-teins. We detected numerous unique membrane proteins in each bacterial isolate, in-cluding ABC transporters and proteins involved in lipopolysaccharide synthesis, when lyophilizing prior to bead-beating compared to only flash freezing prior. As membrane proteins play a central role in AMR resistance mechanisms, this improvement in their isolation and identification will aid in understanding the resistance and molecular mechanisms associated with multi-drug resistant GNB.
HostingRepository	PRIDE
AnnounceDate	2026-05-28
AnnouncementXML	Submission_2026-05-27_22:33:54.802.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Breyer-Lynne Woodland
SpeciesList	scientific name: Escherichia coli; NCBI TaxID: NEWT:562;
ModificationList	carbamoylated residue; S-carboxamidoethyl-L-cysteine; monohydroxylated residue; deamidated residue
Instrument	Q Exactive Plus

Revision	Datetime	Status	ChangeLog Entry
0	2026-03-04 02:44:41	ID requested
1	2026-03-08 15:21:39	announced
⏵ 2	2026-05-27 22:33:55	announced	2026-05-28: Updated project metadata.

Dataset with its publication pending

submitter keyword: proteomics,antimicrobial resistance, membrane proteins, bacterial protein extraction

Matthew P. Padula
contact affiliation	School of Life Sciences and Proteomics and Metabolomics Core Facility, Faculty of Science, University of Technology Sydney, Ultimo, NSW 2007, Australia
contact email	Matthew.Padula@uts.edu.au
lab head
Breyer-Lynne Woodland
contact affiliation	University of Technology Sydney
contact email	breyer-lynne.woodland@student.uts.edu.au
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD075198
     1. Label: PRIDE project
     2. Name: Lyophilization prior to homogenisation and extraction increases membrane protein detection in Gram-negative bacterial proteomic analyses