Global analysis of the Brucella melitensis proteome: Identification of proteins expressed in laboratory-grown culture

Wagner, Mary Ann; Eschenbrenner, Michel; Horn, Troy; Kraycer, Jo Ann; Mujer, Cesar V.; Hagius, Sue D.; Elzer, Philip H.; DelVecchio, Vito G.

doi:10.1002/1615-9861(200208)2:8<1047::aid-prot1047>3.0.co;2-8

Cited by 81 publications

(61 citation statements)

References 19 publications

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“…The first, protein 12, found here in B. suis, was recently also characterized by a proteomic approach in B. melitensis (42). Genes encoding homologues of this protein were found in various organisms, and the highest identity within the putative bacterial proteins was concentrated in a stretch of Ͻ30 amino acids.…”

Section: Discussionmentioning

confidence: 99%

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Release of Periplasmic Proteins of Brucella suis upon Acidic Shock Involves the Outer Membrane Protein Omp25

et al. 2004

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The survival and replication of Brucella in macrophages is initially triggered by a low intraphagosomal pH. In order to identify proteins released by Brucella during this early acidification step, we analyzed Brucella suis conditioned medium at various pH levels. No significant proteins were released at pH 4.0 in minimal medium or citrate buffer, whereas in acetate buffer, B. suis released a substantial amount of soluble proteins. Comparison of 13 N-terminal amino acid sequences determined by Edman degradation with their corresponding genomic sequences revealed that all of these proteins possessed a signal peptide indicative of their periplasmic location. Ten proteins are putative substrate binding proteins, including a homologue of the nopaline binding protein of Agrobacterium tumefaciens. The absence of this homologue in Brucella melitensis was due to the deletion of a 7.7-kb DNA fragment in its genome. We also characterized for the first time a hypothetical 9.8-kDa basic protein composed of five amino acid repeats. In B. suis, this protein contained 9 repeats, while 12 were present in the B. melitensis orthologue. B. suis in acetate buffer depended on neither the virB type IV secretory system nor the omp31 gene product. However, the integrity of the omp25 gene was required for release at acidic pH, while the absence of omp25b or omp25c displayed smaller effects. Together, these results suggest that Omp25 is involved in the membrane permeability of Brucella in acidic medium.Bacteria of the genus Brucella are gram-negative facultative intracellular pathogens of various wild and domestic mammals and are able to cause severe zoonotic infections in humans. Traditionally, three major species are distinguished by their predilections for certain animal hosts: Brucella abortus for cattle, Brucella melitensis for caprines, and Brucella suis for hogs. Whereas B. abortus is the livestock pathogen with the greatest economic impact, B. melitensis and B. suis account for most clinical cases in humans (1, 2, 11).To evade host defenses, Brucella can inhibit neutrophil degranulation and block tumor necrosis factor (TNF) production by macrophages. It has been shown that membrane integrity, in terms of both smooth lipopolysaccharide and outer membrane proteins, is required for such virulent behavior. Furthermore, studies using transposon or signature-tagged mutagenesis have unraveled, with respect to Brucella virulence, the crucial role of an operon homologous to the virB operon of Agrobacterium tumefaciens encoding a type IV secretion system (16, 21, 28). The virulence regulon of A. tumefaciens is triggered in response to chemical signals released at the plant wound site, such as acetosyringone and low pH. Type IV secretion system production is potentiated by monosaccharides (galactose and arabinose) through binding to the periplasmic multiple sugar binding protein ChvE, as well as by low pH (6). However, it was found that under neutral conditions, this secretory system is already produced in B. melitensis or B. abortus, while ...

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Section: Discussionmentioning

confidence: 99%

“…One striking exception was the very basic theoretical pI of 9.44 for the highly charged protein 13. This might explain why its B. melitensis orthologue, unlike that of protein 12, could not be detected by two-dimensional gel electrophoresis proteomic analysis (42).…”

Section: Vol 72 2004 B Suis Releases Periplasmic Proteins At Acidimentioning

confidence: 99%

Release of Periplasmic Proteins of Brucella suis upon Acidic Shock Involves the Outer Membrane Protein Omp25

et al. 2004

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“…Our laboratory has been involved in a comprehensive analysis of the B. melitensis 16M proteome, and initial results have been published recently (40). Previous proteomics studies using B. melitensis cells grown under different conditions have been reported (36,37), and initial work on the B. abortus proteome has been described (26,29).…”

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confidence: 95%

“…Digestion with trypsin was performed in accordance with the ProGest default long trypsin digestion protocol, which was altered to include modification of the tryptic peptides with o-methylisourea (o-MU) (40). Tryptic peptides were modified for 1 h at 37°C with 1 M o-MU in 100 mM ammonium bicarbonate at pH 10.0 as previously described (24).…”

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confidence: 99%

Comparative Proteome Analysis ofBrucella melitensisVaccine Strain Rev 1 and a Virulent Strain, 16M

et al. 2002

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The genus Brucella consists of bacterial pathogens that cause brucellosis, a major zoonotic disease characterized by undulant fever and neurological disorders in humans. Among the different Brucella species, Brucella melitensis is considered the most virulent. Despite successful use in animals, the vaccine strains remain infectious for humans. To understand the mechanism of virulence in B. melitensis, the proteome of vaccine strain Rev 1 was analyzed by two-dimensional gel electrophoresis and compared to that of virulent strain 16M. The two strains were grown under identical laboratory conditions. Computer-assisted analysis of the two B. melitensis proteomes revealed proteins expressed in either 16M or Rev 1, as well as up-or down-regulation of proteins specific for each of these strains. These proteins were identified by peptide mass fingerprinting. It was found that certain metabolic pathways may be deregulated in Rev 1. Expression of an immunogenic 31-kDa outer membrane protein, proteins utilized for iron acquisition, and those that play a role in sugar binding, lipid degradation, and amino acid binding was altered in Rev 1.Brucellosis is a major infectious disease afflicting humans and a wide range of domesticated animals worldwide. The disease is caused by several Brucella species, which are aerobic, nonmotile, gram-negative, facultative intracellular coccobacilli. The genus Brucella belongs to the ␣-2 subgroup of the class Proteobacteria. It is subdivided, on the basis of its pathogenicity and host preference, into six nomen species: Brucella abortus, B. canis, B. melitensis, B. neotomae, B. ovis, and B. suis (12). In addition, a new strain affecting marine mammals was recently isolated and tentatively named B. maris (32). On the basis of DNA hybridization data, it was suggested that all of these organisms be placed into a single species, B. melitensis (39). Among the various nomen species, B. abortus, B. canis, B. suis, B. maris, and B. melitensis have been reported to infect humans (21,35,13). B. melitensis is a pathogen of goats and sheep and is considered the most virulent species for humans. Human infection can result from either occupational contact or ingestion of contaminated food.Vaccination and eradication of infected hosts have been key factors in the control of brucellosis. Rev 1, an attenuated strain of virulent B. melitensis, was developed in 1957 (19). It is considered the most effective vaccine for the control of brucellosis in small ruminants and was used in comprehensive vaccination programs in many countries, including Saudi Arabia, Kuwait, Mongolia, Spain, and Turkey (8).Our laboratory has been involved in a comprehensive analysis of the B. melitensis 16M proteome, and initial results have been published recently (40). Previous proteomics studies using B. melitensis cells grown under different conditions have been reported (36, 37), and initial work on the B. abortus proteome has been described (26, 29). A comparative study was conducted with B. abortus vaccine strains S19 and RB51 and...

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“…The central role of this enzyme in the synthesis of hexoses derived from glucose makes pleiotropic effects on the synthesis of oligo-and polysaccharides likely and, at least, the pgm mutant is also blocked in the synthesisof the periplasmic (1,2) cyclic glucans . Mutation in the homologous gene of B. melitensis causes a core defect as judged by the electrophoretic mobility in SDS-PAGE of its R-LPS (30,87,88)). …”

Section: B Abortus B2211 Pgmmentioning

confidence: 99%

Brucellosis Vaccines: An Overview

Reza¹,

Kazemi²,

Fatemeh³

et al. 2012

Zoonosis

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Global analysis of the Brucella melitensis proteome: Identification of proteins expressed in laboratory-grown culture

Cited by 81 publications

References 19 publications

Release of Periplasmic Proteins of Brucella suis upon Acidic Shock Involves the Outer Membrane Protein Omp25

Release of Periplasmic Proteins of Brucella suis upon Acidic Shock Involves the Outer Membrane Protein Omp25

Comparative Proteome Analysis ofBrucella melitensisVaccine Strain Rev 1 and a Virulent Strain, 16M

Brucellosis Vaccines: An Overview

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