Ultrastructural and glycoproteomic characterization of Prevotella intermedia: Insights into O ‐glycosylation and outer membrane vesicles

Electron cryotomography analysis revealed ultrastructural details ofPrevotella intermedia, revealing an electron-dense surface layer surrounding both cells and outer membrane vesicles (OMVs). Mass spectrometry analysis enabled the identification of a single majorO-glycan [dHex-dHex-HexNAc(HPO3-C6H12O5)-dHex-Hex-HexA-Hex(dHex)] found in 443 unique sites within 224 glycoproteins. Bioinformatic analyses ofO-glycoprotein localization predicted 73 periplasmic proteins, 53 inner membrane proteins, 52 lipoproteins, 26 outer membrane proteins, and 14 proteins secreted by the type IX secretion system. AbstractPrevotella intermedia, a Gram-negative bacterium from the Bacteroidota phylum, is associated with periodontitis. Other species within this phylum are known to possess the generalO-glycosylation system. TheO-glycoproteome has been characterized in several species, includingTannerella forsythia,Porphyromonas gingivalis, andFlavobacterium johnsoniae. In our study, we used electron cryotomography (cryoET) and glycoproteomics to reveal the ultrastructure ofP. intermedia and characterize itsO-glycoproteome. Our cryoET analysis unveiled the ultrastructural details of the cell envelope and outer membrane vesicles (OMVs) ofP. intermedia. We observed an electron-dense surface layer surrounding both cells and OMVs. The OMVs were often large (>200  nm) and presented two types, with lumens being either electron-dense or translucent. LC-MS/MS analyses ofP. intermedia fractions led to the ...

Source: MicrobiologyOpen - February 27, 2024 Category: Microbiology Authors: Xi Ye, Bindusmita Paul, Joyce Mo, Eric C. Reynolds, Debnath Ghosal, Paul D. Veith Tags: COMMENTARY Source Type: research