Proteomic shifts in multi-species oral biofilms caused by Anaeroglobus geminatus

Abstract

Anaeroglobus geminatus is a relatively newly discovered putative pathogen, with a potential role in the microbial shift associated with periodontitis, a disease that causes inflammatory destruction of the periodontal tissues, and eventually tooth loss. This study aimed to introduce A. geminatus into a polymicrobial biofilm model of relevance to periodontitis, and monitor the proteomic responses exerted to the rest of the biofilm community. A. geminatus was grown together with another 10-species in a well-established “subgingival” in vitro biofilm model. Its effects on the other species were quantitatively evaluated by qPCR and label-free proteomics. A. geminatus caused a significant increase in P. intermedia numbers, but not the other species in the biofilm. Whole cell proteome profiling of the biofilms by LC-MS/MS identified a total of 3213 proteins. Label-free quantitative proteomics revealed that 187 proteins belonging to the other 10 species were differentially abundant when A. geminatus was present in the biofilm. The species with most up-regulated and down-regulated proteins were P. intermedia and S. oralis, respectively. Regulated proteins were of primarily of ribosomal origin, and other affected categories involved proteolysis, carbon metabolism and iron transport. In conclusion, A. geminatus can be successfully grown in a polymicrobial biofilm community, causing quantitative proteomic shifts commensurate with increased virulence properties.

Host‐microbe interactions: Profiles in the transcriptome, the proteome, and the metabolome

“…Many in vitro proteomic studies related to periodontal disease and periodontal pathogens have been carried out in the last decade. [66][67][68][69][70][71][72][73][74][75][76][77][78][79][80][81] Of the oral pathogens examined, 81 the two most commonly investigated bacterial species are Aggregatibacter actinomycetemcomitans and Porphyromonas gingivalis. 66,67,69,70,73,[76][77][78]80 Porphyromonas gingivalis, a major pathogen in periodontitis, releases virulence factors, such as gingipains, through outer membrane vesicles 66 via a C-terminal domain signal system.…”

Streptococcus oralis maintains homeostasis in oral biofilms by antagonizing the cariogenic pathogen Streptococcus mutans

“…Deciphering the interactions between species within biofilms is crucial for understanding the factors that ensure their stability, or those that drive changes that lead to dysbiosis. The Zürich in vitro biofilm models are useful tools to define such interactions of groups of species, individual species, or their virulence factors …”

Pressure Cycling Technology Assisted Mass Spectrometric Quantification of Gingival Tissue Reveals Proteome Dynamics during the Initiation and Progression of Inflammatory Periodontal Disease

“…Their concentrations were measured using a NanoDrop 1000 spectrophotometer (Thermo Fisher), and adjusted to 0.3 µg µL −1 . 0.6 µg (2 µL) of peptides were injected into an Orbitrap Fusion MS (Thermo Fisher Scientific) for proteomic analysis as described previously (Figure 1B). Briefly, peptides were separated using a Thermo Scientific EASY‐nLC 1200 system (Thermo Fisher Scientific) coupled to a 15 cm‐long and 75 µm‐diameter silica emitter, packed with a ReproSil‐Pur C18‐AQ 120 A and 1.9 µm resin (Dr. Maisch HPLC GmbH).…”

“…The protein label‐free quantification (LFQ) was performed by Progenesis QI for proteomics software (Nonlinear Dynamics) as descript previously . In brief, the raw files of each sample were aligned with a pooled sample for peak picking and creating Mascot files (.mgf).…”