PXD056875-1

PXD056875 is an original dataset announced via ProteomeXchange.

Title	Uncovering new insights into bacterial pseudaminylation with a pan-specific antibody tool
Description	Pseudaminic acids (Pse) are a family of nine-carbon carbohydrates produced by many bacteria, including important human pathogens. These non-mammalian carbohydrates are found within the structure of lipopolysaccharides, capsular polysaccharides, and as O-linked post-translational modifications (PTMs) of serine and threonine residues within flagellin and pili proteins. This protein O-pseudaminylation is important for the host colonisation, virulence and motility of pathogens, although only a handful of cell surface proteins bearing this PTM have been identified to date, and the functional role of this PTM remains largely unknown. Herein, we describe the synthesis of O-pseudaminylated serine and threonine amino acids with both α- and β-anomeric configuration and their installation into tryptic glycopeptide fragments of Helicobacter pylori flagellin A and B. These synthetic glycopeptides enabled assignment of the stereochemistry of O-linked Pse in H. pylori and facilitated the generation of monoclonal antibodies (mAbs) that can recognise both α- and β-configured Pse (and its C8 epimer) linked to protein or glycan. The sequences, affinities and structural epitopes of these pan-specific Pse mAbs were determined, informing the development of an immunoenrichment strategy for glycoproteomic analyses that expanded the number of known pseudaminylation sites and substrates across the proteomes of pathogenic Helicobacter pylori and Campylobacter jejuni strains. These mAbs are also capable of enriching the more complex O-glycans that contain terminal Pse in pathogenic Acinetobacter baumannii strains, enabling the discovery of hitherto unknown glycoproteins. Finally, we demonstrate that these mAbs recognise a variety of different multidrug resistant Acinetobacter baumannii capsule types and are capable of clearing infections in mice.
HostingRepository	PRIDE
AnnounceDate	2025-10-25
AnnouncementXML	Submission_2025-10-25_00:31:10.310.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Mark Larance
SpeciesList	scientific name: Helicobacter pylori P12; NCBI TaxID: NEWT:570508;
ModificationList	acetylated residue; monohydroxylated residue; iodoacetamide derivatized residue
Instrument	Orbitrap Fusion Lumos

Revision	Datetime	Status	ChangeLog Entry
0	2024-10-16 16:28:16	ID requested
⏵ 1	2025-10-25 00:31:11	announced

Dataset with its publication pending

submitter keyword: Pseudaminic acid,glycosylation, bacteria, flagella

Mark Larance
contact affiliation	Charles Perkins Centre, School of Medical Sciences, University of Sydney, Sydney, Australia.
contact email	mark.larance@sydney.edu.au
lab head
Mark Larance
contact affiliation	The University of Sydney
contact email	mark.larance@sydney.edu.au
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD056875
     1. Label: PRIDE project
     2. Name: Uncovering new insights into bacterial pseudaminylation with a pan-specific antibody tool