Dataset Information

Characterization of the O-Glycoproteome of Flavobacterium johnsoniae

ABSTRACT: Flavobacterium johnsoniae is a free-living member of the Bacteroidota phylum found in soil and water. It is frequently used as a model species for studying a type of gliding motility dependent on the type IX secretion system (T9SS). O-glycosylation has been reported in several Bacteroidota species and the O-glycosylation of S-layer proteins in Tannerella forsythia was shown to be important for certain virulence features. In this study we characterised the O-glycoproteome of F. johnsoniae and identified 325 O-glycosylation sites within 226 glycoproteins. The structure of the major glycan was found to be a hexasaccharide with the sequence Hex–(Me-dHex)–Me-HexA–Pent–HexA–Me-HexNAcA. Bioinformatic localisation of the glycoproteins determined 68 inner membrane proteins, 60 periplasmic proteins, 26 outer membrane proteins, 57 lipoproteins and 9 proteins secreted by the T9SS. The glycosylated sites were predominantly located in the periplasm where they are postulated to be beneficial for protein folding/stability. Six proteins associated with gliding motility or the T9SS were demonstrated to be O-glycosylated.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Flavobacterium Johnsoniae Uw101

SUBMITTER: Paul Veith

LAB HEAD: Eric C Reynolds

PROVIDER: PXD040877 | Pride | 2023-04-18

REPOSITORIES: Pride

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			Action	DRS
	210930_PAul_PV-3097_CE1.raw	Raw
	210930_PAul_PV-3097_CE10.raw	Raw
	210930_PAul_PV-3097_CE11.raw	Raw
	210930_PAul_PV-3097_CE12.raw	Raw
	210930_PAul_PV-3097_CE2.raw	Raw

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Project description:N-glycosylation is an important posttranslational modification in all eukaryotes, but little is known about the N-glycoproteomes in microalgal systems. Here, N-glycoproteome of the model diatom Phaeodactylum tricornutum was unveiled. Totally, 893 different N-glycosylated sites from 649 proteins were identified from P. tricornutum. The new synthesized N-glycans were principally transferred to asparagine residues within the conserved N-X-S/T (where X is a residue other than proline) sequence of nascent polypeptide chains. Functional annotation of N-glycosylated proteins showed that 70% N-glycoproteins were involved in vesicular transport and posttranslational modification, protein turnover, chaperones, indicating the N-glycosylation was important for these functions. Of the all identified N-glycoproteins 45.3% were predicted to localize on chloroplast, which implied the widespread regulatory role of N-glycosylation in chloroplast. Furthermore, the enrichment results of N-glycoproteins indicated that compared to ‘Cellular component’ and ‘Biological process’ categories proteins related with the ‘Molecular function’ category were more prone to be N-glycosylated. And it was speculated that N-glycosylation played a vital regulatory role in catalytic activity of enzymes and metabolism processes of many small molecules. 47% of all enriched proteins were related with metabolic pathways. The functional annotation and enrichment of N-glycoproteins suggested that N-glycoproteins participated in a variety of important metabolic pathways and perform different functions in P. tricornutum. 12 proteins involved in the ER quality control mechanism and ER- associated degradation pathway were identified as N-glycosylated proteins, indicating that the N-glycosylated modification was important for their functions in the protein N-glycosylation pathway. Additionally, some interacted glycoproteins were classified from this study, which provided valuable information for studying the functions of these glycoproteins. In the study, the identification of N-glycosylation on nascent proteins expands our understanding of this PTM at a proteomics scale and may facilitate the elucidation of the precise function of proteins in this model diatom.

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Characterization of the O-Glycoproteome of Flavobacterium johnsoniae

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