Dataset Information

The proteome of Rickettsia akari revealed a 44kDa-OMP as novel biomarker for Rickettsialpox

ABSTRACT: Our study aimed to investigate the proteome structure of R. akari and unveil immunoreactive surface-exposed proteins using the latest proteomics techniques. To achieve these goals, we applied the Shotgun protein identification approach; membrane surface protein enrichment methods combined with liquid chromatography-tandem mass spectrometry (LC-MS/MS) and immunoblotting techniques. We identified aroun 300 unique proteins from the whole-cell extract exhibiting that the most represented proteins were those involved in translation, post-translational modifications, energy production, and cell wall development. A significant part belongs to amino acid transport and intracellular trafficking. Besides, several proteins affecting virulence were detected. Then, in silico analysis from detected surface proteins revealed that 25 putative predicted outer membrane proteins contain beta-barrel structure and 11 proteins were predicted to have secretion signal peptide sequence.

INSTRUMENT(S): LTQ Orbitrap

ORGANISM(S): Rickettsia Akari

TISSUE(S): Bacterial Protein, Vero Cell, Egg

DISEASE(S): Rickettsialpox

SUBMITTER: Jiri DRESLER

LAB HEAD: Jiri Dresler

PROVIDER: PXD017762 | Pride | 2020-07-13

REPOSITORIES: Pride

ACCESS DATA

Dataset's files

Source:

			Action	DRS
	09092019_29_AF_2019_05_EDA_JD2_R.akariculturedoneggs4mgmL.raw	Raw
	09092019_30_AF_2019_05_EDA_JD3VEROcelllineinfectedRakari.raw	Raw
	09092019_32_AF_2019_05_EDA_JD4R.akariculturedoneggsinALTlysisbuffer.raw	Raw
	Databases_and_annotations.zip	Other
	MaxQuant_results.zip	Other

Items per page:

1 - 5 of 6

Publications

Proteomic analysis of Rickettsia akari proposes a 44 kDa-OMP as a potential biomarker for Rickettsialpox diagnosis.

Csicsay František F Flores-Ramirez Gabriela G Zuñiga-Navarrete Fernando F Bartošová Mária M Fučíková Alena A Pajer Petr P Dresler Jiří J Škultéty Ľudovít Ľ Quevedo-Diaz Marco M

BMC microbiology 20200708 1

<h4>Background</h4>Rickettsialpox is a febrile illness caused by the mite-borne pathogen Rickettsia akari. Several cases of this disease are reported worldwide annually. Nevertheless, the relationship between the immunogenicity of R. akari and disease development is still poorly understood. Thus, misdiagnosis is frequent. Our study is aiming to identify immunogenic proteins that may improve disease recognition and enhance subsequent treatment. To achieve this goal, two proteomics methodologies w ...[more]

PMID: 32640994

Similar Datasets

Project description:Obesity-induced insulin resistance of the liver is characterised by increased gluconeogenesis, which contributes to elevated blood glucose levels in individuals with type 2 diabetes. Research into how fatty acids induce insulin resistance has commonly focused on the induction of insulin resistance. We hypothesise that by shifting focus to the reversal of an insulin resistant phenotype, novel insights can be made into the mechanisms by which insulin resistance can be overcome. Using global gene and lipid expression profiling, we aimed to identify biological pathways altered in parallel with restoration of palmitate-induced deregulation of glucose production using metformin and sodium salicylate. FAO hepatoma cells were treated with palmitate (0.075mM, 48h) with or without metformin (0.25mM) and sodium salicylate (2mM) in the final 24h of palmitate treatment, and effects on glucose production were determined. Microarray followed by gene set enrichment analysis was performed to investigate pathway regulation. A lipidomic analysis (HPLC-MS/MS) and measurement of secreted bile acids and cholesterol were performed. Reversal of palmitate-induced impairment of glucose production by metformin and sodium salicylate was characterised by down-regulated expression of metabolic pathways regulating acetyl-CoA to cholesterol and bile acid biosynthesis. Total levels of intracellular and secreted cholesterol and bile acids were not different between impaired and restored glucose production. Total intracellular levels of diacylgycerol, triacylglycerol and cholesterol esters increased with palmitate (impaired glucose production), however, these were not further altered with metformin and sodium salicylate (restored glucose production). Six individual lipid species containing 18:0 and 18:1 side-chains were reduced by metformin and sodium salicylate. Widespread lipid metabolism changes induced by the reversal of palmitate-induced deregulation of glucose production with metformin and sodium salicylate were identified. While cholesterol and bile acid levels remained unchanged, the flux through these pathways may in part explain these findings. The identification of lipid species containing 18:0 and 18:1 side chains being regulated alongside changes to glucose production may indicate potential mediators of glucose production and insulin resistance. Three-condition experiment, Vehicle, Palmitate (PA) and Palmitate (PA) + Metformin (Met) + Sodium Sailcylate (NaS) with biological replicates: 8 Vehicle, 20 PA and 20 PA+Met+NaS , independently grown and harvested. One replicate per array.

Project description:Specific Lactobacillus strains are marketed as probiotics i.e. health promoting organisms. As previously shown in vivo, L. plantarum WCFS1 modulates immune responses via nuclear factor (NF)kB in a growth phase dependent manner, likely caused by changed microbial cell surface characteristics. Here, we identified the differential cell-surface proteome composition of L. plantarum WCFS1 cells derived from the mid-logarithmic and late stationary growth phase by surface trypsination of intact bacterial cells, followed by a combined 1D- and 2D-LC-MS shotgun proteomics strategy, and growth phase dependent transcriptome analysis. Overall, these analyses led to the identification of 31 surface proteins differentially present in the logarithmic and stationary phase, whereas additional 17 and 33 proteins appeared solely present in stationary and logarithmic phase of growth, respectively. From 75% of predicted surface related genes displaying relatively high expression levels, the corresponding proteins were detected, demonstrating a high correlation between transcriptome and proteome profiles. The impact of cell-surface peptide fractions on cellular host responses was evaluated using NFkB promoter activation assays in intestinal epithelial cells, revealing that only late stationary phase derived peptides are capable of attenuating flagellin and cell envelope induced NFkB activation. Overall our data mechanistically expand earlier observations that revealed growth-phase dependent immunomodulation by L. plantarum WCFS1, leading to a typical adjuvant- or tolerance-like response after exposure to stationary phase harvested bacterial cells. Sub-fractionation of the late stationary phase peptide fraction revealed a sub-set of strong attenuator peptides able to manipulate NFkB related pathways to hereby attenuate pro-inflammatory signaling. Samples were hybridizedin a loop design linking each timepoint to the next as well as the previous timepoint. Additional hybridizations were introduced linking each sample three timepoints forward and three timepoints back. All samples were hybridized unsing at least each of the two dyes once.