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Abstract
Brucellosis is a zoonotic infection caused by bacteria of the genus Brucella. The species, B. abortus and B. melitensis, major causative agents of human brucellosis, share remarkably similar genomes, but they differ in their natural hosts, phenotype, antigenic, immunogenic, proteomic and metabolomic properties. In the present study, label-free quantitative proteomic analysis was applied to investigate protein expression level differences. Type strains and field strains were each cultured six times, cells were harvested at a midlogarithmic growth phase and proteins were extracted. Following trypsin digestion, the peptides were desalted, separated by reverse-phase nanoLC, ionized using electrospray ionization and transferred into an linear trap quadrapole (LTQ) Orbitrap Velos mass spectrometer to record full scan MS spectra (m/z 300–1700) and tandem mass spectrometry (MS/MS) spectra of the 20 most intense ions. Database matching with the reference proteomes resulted in the identification of 826 proteins. The Cluster of Gene Ontologies of the identified proteins revealed differences in bimolecular transport and protein synthesis mechanisms between these two strains. Among several other proteins, antifreeze proteins, Omp10, superoxide dismutase and 30S ribosomal protein S14 were predicted as potential virulence factors among the proteins differentially expressed. All mass spectrometry data are available via ProteomeXchange with identifier PXD006348.
Highlights
Brucella represents a Gram-negative bacterial genus of the α-2 subgroup of Proteobacteria
Of the 826 proteins identified, 102 proteins (12%) were predicted to be potentially virulence-associated (Supplementary Table 3) among which 22 proteins were identified as differentially expressed proteins (16 upregulated and six downregulated in B. abortus comparison to B. melitensis)
Our quantitative proteomic analysis of reference and field-isolated strains of B. abortus and B. melitensis not surprisingly confirms the existence of proteome level differences between the strains
Summary
Brucella represents a Gram-negative bacterial genus of the α-2 subgroup of Proteobacteria. Brucellae are highly adapted to their intracellular lifestyle and are the causative agents of human and animal brucellosis (“undulant fever”, “Malta fever”, “Mediterranean fever” or “Bang’s disease”) [1]. They are highly infective and 10–100 bacteria cause human infection [2,3]. The classification of Brucella species is under debate due to the reported high degree of homology found in DNA-DNA hybridization studies. This results in the proposal that the genus Brucella is a single genomospecies and the species are only biovars of B. melitensis [5,6]. The Centers for Disease Control and Prevention has listed Brucella species on the Federal Select Agent Program/Select Agents and Toxins List (https://www.selectagents.gov/SelectAgentsandToxinsList.html, assessed in March 2020) and on the Emergency Preparedness and Response Bioterrorism Agents/Diseases List (https://emergency.cdc.gov/agent/agentlist.asp, assessed in March 2020) as prepared by the National Center for Emerging and Zoonotic Infectious Diseases
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