Global analysis of lysine succinylome in the periodontal pathogen Porphyromonas gingivalis

Abstract

The gram-negative anaerobe Porphyromonas gingivalis is not only a keystone periodontal pathogen but also an emerging systemic pathogen. Although the newly discovered protein post-translational modification (PTM), lysine succinylation (Ksuc), appears to play an important role in modulating metabolic processes in bacteria, this PTM has not been investigated in P gingivalis. In this study, we used a highly sensitive proteomics approach combining affinity enrichment with high-resolution liquid chromatography coupled with tandem mass spectrometry to examine Ksuc in P gingivalis.In total, 345 Ksuc sites in 233 proteins were identified and determined to be involved in a variety of cellular processes. In the region surrounding Ksuc sites, lysine residues were drastically overrepresented and sequence motifs with succinyl-lysine flanked by a lysine at the +3 or +6 positions appear to be unique to this pathogen. Additionally, our results suggest a crosstalk between Ksuc and glycosylation, but the overlap between Ksuc and acetylation in P gingivalis is quite different from that observed in other organisms. Notably, Ksuc was observed in proteins associated with established virulence factors, including gingipains, fimbriae, RagB, and PorR. Moreover, products of the factors necessary for P gingivalis in vitro survival (18.5%) were found to be succinylated at lysine sites and the same was observed in products of fitness factors for P gingivalis survival in both abscess and epithelial cell colonization environments (12%). Collectively, these results suggest that Ksuc may be a new mechanism in modulating the virulence, adaptation, and fitness of P gingivalis. K E Y W O R D S fitness factors, lysine succinylation, necessary factors, Porphyromonas gingivalis, proteomics | 75 WU et al.

Comprehensive profiling of protein lysine acetylation and its overlap with lysine succinylation in the Porphyromonas gingivalis fimbriated strain ATCC 33277

“…Moreover, the GO functional classification and enrichment results for acetylated proteins in the present study were highly similar to that for succinylated proteins (Wu et al., 2019). Notably, enzymes associated with metabolism represented the largest group of both acetylated proteins and succinylated proteins.…”

“…et al have identified 130 Kac sites from 92 proteins in P. gingivalis W50 (Butler et al, 2015). Recently, we have identified 345 Ksuc sites in 233 proteins in P. gingivalis ATCC 33277 (Wu et al, 2019), supporting a possible regulatory role of Kac and Ksuc in P. gingivalis. As previously reported, we further analyzed the overlap between these two PTMs in P. gingivalis (Wu et al, 2019).…”

“…Recently, we have identified 345 Ksuc sites in 233 proteins in P. gingivalis ATCC 33277 (Wu et al., 2019), supporting a possible regulatory role of Kac and Ksuc in P. gingivalis . As previously reported, we further analyzed the overlap between these two PTMs in P. gingivalis (Wu et al., 2019). However, the comparison showed that the overlap between Ksuc and Kac is less extensive than that in other organisms (Gaviard et al., 2018; Weinert et al., 2013).…”

“…(b) Venn diagram showing the number of peptides with Ksuc modification, Kac modification, and both modifications. (c) Example of overlap between Ksuc and Kac: Catalytic enzymes involved in a proposed metabolic map of energy and cytotoxin production in P. gingivalis ATCC 33277 (Hendrickson et al., 2009; Sato et al., 2016; Takahashi et al., 2000; Wu et al., 2019). Catalytic enzymes were represented by their PGN designation; proteins with Ksuc modification were labeled with a red star, the ones with Kac modification were labeled with a green star and the ones with both modifications were labeled with a yellow star…”

“…However, our previous study found that the overlap between Ksuc (Hendrickson et al, 2009;Sato et al, 2016;Takahashi et al, 2000;Wu et al, 2019). Catalytic enzymes were represented by their PGN designation; proteins with Ksuc modification were labeled with a red star, the ones with Kac modification were labeled with a green star and the ones with both modifications were labeled with a yellow star this difference may be due to the different P. gingivalis strains used, we compared the previous lysine succinylome data and present lysine acetylome data both in P. gingivalis ATCC 33277.…”