Proteomics ofPseudomonas aeruginosa: the increasing role of post-translational modifications

Abstract

Pseudomonas aeruginosa is a Gram-negative opportunistic pathogen widely involved in human infections. The high occurrence of this bacterial species in the clinical field is due to its high ability to adapt to detrimental environments, in particular its strong inherent antibiotic resistance, its ability to form biofilms and to produce virulence factors. The application of proteomics to clinical microbiology is probably one of the most innovative strategies of the last decades to understand complex microbial systems, by providing individual proteome charts of pathogens. Areas covered: In the last decade, proteomic advances have allowed in high-throughput the screening of proteins modified by diverse co- and post-translational modifications in P. aeruginosa. This review will present the current state of the art for the characterization of PTMs in P. aeruginosa by proteomics approaches. We will then discuss on the involvement of PTMs in P. aeruginosa physiology. Expert commentary: Modified proteins and enzymes involved in the addition/removal of modifications will surely constitute targets of interest to develop new therapeutic drugs to fight against P. aeruginosa.

Exoproteomics for Better Understanding Pseudomonas aeruginosa Virulence

“…Virulence factors are taken by chaperones for their export and investigating chaperones partners can be informative for the discovery of new effectors [16,39]. Components of T3SS and T6SS export apparatus were also observed with different PTMs [50]. Without N-glycosylation, the virulence factor LecB cannot be exported in P. aeruginosa periplasm [117].…”

“…Next, the bottom-up strategy (or shotgun) is mainly used for the characterisation of exoproteins in bacteria. Recent reviews have been published describing the different available proteomic approaches for protein identification [50][51][52]. Briefly, proteins are digested with a specific endoprotease (generally trypsin) to yield peptides that will be analysed by high-sensitive and high-resolution nanoLC-MS/MS.…”

“…Moreover, 2-DE and SDS-PAGE have the great benefit to allow the visualisation of proteins and the determination of protein abundance by gel comparisons using software packages. Another advantage for 2-DE is the ability to distinguish spots with different isoelectric points (pI) and/or molecular weights (MW) for a same protein, suggesting the presence of PTMs [50]. Due to the presence of proteases in bacterial supernatants, a protein may be present at different MWs.…”

“…Nowadays, proteomics is the tool of choice to identify, quickly and reliably, diverse modifications ( Figure 2). In the past two decades, PTMs have been successfully characterised in intracellular medium by different proteomic strategies previously reviewed [50,85]. In P. aeruginosa, the studies of phosphorylation [63,86], lysine acetylation [87,88], lysine succinylation [88], or glycosylation [89] have been performed for intracellular media revealing PTMs on proteins involved in virulence.…”

There is no hiding if you Seq: recent breakthroughs in Pseudomonas aeruginosa research revealed by genomic and transcriptomic next-generation sequencing

“…Indeed, Pseudomonas sp. post-translationally modify many classes of enzymes and structural proteins in many different ways (reviewed in Gaviard et al (35)) and it is probable that many of these will be significant for the general physiology of the bacterium and/or for virulence. Proteomic analysis is not only likely to validate predictions generated from transcriptome data, but also to answer questions that NGS approaches cannot currently address.…”