Dataset Information

O-glycoproteome of Porphyromonas gingivalis

ABSTRACT: Porphyromonas gingivalis is an important human pathogen and also a model organism for the Bacteroidetes phylum. O-glycosylation has been reported in this phylum with findings that include the O-glycosylation motif, the structure of the O-glycans in a few species and an extensive O-glycoproteome analysis in Tannerella forsythia. However, O-glycosylation has not yet been confirmed in P. gingivalis. We therefore used glycoproteomics approaches including partial deglycosylation with trifluoromethanesulfonic acid as well as both HILIC and FAIMS based glycopeptide enrichment strategies leading to the identification of 257 putative glycosylation sites in 145 glycoproteins. The sequence of the major O-glycan was elucidated to be Hex-(dHex)-HexA-Hex-dHex-HexNAc-(P-Gro-[Ac]0-2)-HexNAc. Western blot analyses of mutants lacking the glycosyltransferases PGN_1134 and PGN_1135 demonstrated their involvement in the biosynthesis of the glycan while mass spectrometry analysis of the truncated O-glycans suggested that PGN_1134 and PGN_1135 transfer the two HexNAc sugars. Interestingly, a strong bias against the O-glycosylation of abundant proteins exposed to the cell surface such as abundant T9SS cargo proteins, surface lipoproteins and outer membrane β-barrel proteins was observed. In contrast, the great majority of proteins associated with the inner membrane or periplasm were glycosylated irrespective of their abundance. The P. gingivalis O-glycosylation system may therefore function to establish the desired physicochemical properties of the periplasm.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Porphyromonas Gingivalis Atcc 33277

DISEASE(S): Periodontitis

SUBMITTER: Paul Veith

LAB HEAD: Eric C Reynolds

PROVIDER: PXD028120 | Pride | 2022-02-15

REPOSITORIES: Pride

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Dataset's files

Source:

			Action	DRS
	200910_PaulV_PV-1115_M01.raw	Raw
	200910_PaulV_PV-1115_M02.raw	Raw
	200910_PaulV_PV-1115_M03.raw	Raw
	200910_PaulV_PV-1115_M04.raw	Raw
	200910_PaulV_PV-1115_M05.raw	Raw

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Publications

Characterization of the O-Glycoproteome of Porphyromonas gingivalis.

Veith Paul D PD Shoji Mikio M Scott Nichollas E NE Reynolds Eric C EC

Microbiology spectrum 20220105 1

Porphyromonas gingivalis is an important human pathogen and also a model organism for the Bacteroidetes phylum. O-glycosylation has been reported in this phylum with findings that include the O-glycosylation motif, the structure of the O-glycans in a few species, and an extensive O-glycoproteome analysis in Tannerella forsythia. However, O-glycosylation has not yet been confirmed in P. gingivalis. We therefore used glycoproteomics approaches including partial deglycosylation with trifluoromethan ...[more]

PMID: 34985300

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