scholarly journals Proteome Analysis of Outer Membrane Vesicles From a Highly Virulent Strain of Haemophilus parasuis

2021 ◽  
Vol 8 ◽  
Author(s):  
Kunli Zhang ◽  
Pinpin Chu ◽  
Shuai Song ◽  
Dongxia Yang ◽  
Zhibiao Bian ◽  
...  
Keyword(s):  
Outer Membrane ◽  
Virulent Strain ◽  
Bacterial Pathogen ◽  
Proteome Analysis ◽  
Membrane Vesicles ◽  
Subcellular Location ◽  
Outer Membrane Vesicles ◽  
Haemophilus Parasuis ◽  
Extracellular Milieu ◽  
Virulence Potential

Haemophilus parasuis has emerged as an important bacterial pathogen in pig husbandry, as H. parasuis can coinfect pigs with a variety of pathogenic microorganisms and further cause an aggravation of the disease. It is crucial to investigate its pathogenetic mechanism. Gram-negative bacteria naturally secrete outer membrane vesicles (OMVs), and their potent virulence factors play prominent roles that affect the interaction between bacteria and host. Still, the pathogenesis that is associated with the bacterial OMVs has not been well-elucidated. In this study, we investigated the secretion of OMVs from a clinical H. parasuis isolate strain (H45). In addition, we further analyzed the characterization, the comprehensive proteome, and the virulence potential of OMVs. Our data demonstrated that H. parasuis could secrete OMVs into the extracellular milieu during infection. Using liquid chromatography with tandem mass spectrometry (MS/MS) identification and bio-information analysis, we identified 588 different proteins associated with OMVs. Also, we also analyzed the subcellular location and biological function of those proteins. These proteins are mainly involved in immune and iron metabolism. Moreover, we confirmed the pathogenicity of H. parasuis OMVs by observing a strong inflammatory response in J774A.1 and porcine alveolar macrophages. Taken together, our findings suggested that OMVs from H. parasuis were involved in the pathogenesis of this bacterium during infection.

In vitro study of virulence potential of Acinetobacter baumannii outer membrane vesicles

2017 ◽  
Vol 111 ◽  
pp. 218-224 ◽  
Author(s):  
Chandana Jha ◽  
Sujata Ghosh ◽  
Vikas Gautam ◽  
Pankaj Malhotra ◽  
Pallab Ray
Keyword(s):  
Acinetobacter Baumannii ◽  
Outer Membrane ◽  
In Vitro Study ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles ◽  
Vitro Study ◽  
Virulence Potential

scholarly journals Proteome analysis of outer membrane vesicles from a clinicalAcinetobacter baumanniiisolate

2009 ◽  
Vol 297 (2) ◽  
pp. 150-156 ◽  
Author(s):  
Sang-Oh Kwon ◽  
Yong Song Gho ◽  
Je Chul Lee ◽  
Seung Il Kim
Keyword(s):  
Outer Membrane ◽  
Proteome Analysis ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles

scholarly journals The Xanthomonas Ax21 protein is processed by the general secretory system and is secreted in association with outer membrane vesicles

2013 ◽  
Author(s):  
Ofir Bahar ◽  
Rory N Pruitt ◽  
Dee Dee Luu ◽  
Benjamin Schwessinger ◽  
Randy Ruan ◽  
...  
Keyword(s):  
Outer Membrane ◽  
Bacterial Pathogen ◽  
Membrane Vesicles ◽  
Secretion System ◽  
Outer Membrane Vesicles ◽  
Type I ◽  
Broad Spectrum Resistance ◽  
Sec System ◽  
Multiple Strains ◽  
Secretory System

Pattern recognition receptors (PRRs) play an important role in detecting invading pathogens and mounting a robust defense response to restrict infection. In rice, one of the best characterized PRRs is XA21, a leucine rich repeat receptor-like kinase that confers broad-spectrum resistance to multiple strains of the bacterial pathogen Xanthomonas oryzae pv. oryzae (Xoo). In 2009 we reported that an Xoo protein, called Ax21, was secreted by a type I-secretion system and that it served to a ctivate X A 21 -mediated immunity. This report has recently been retracted. Here we present data that corrects our previous model. We first show that Ax21 secretion does not depend on the predicted type I secretion system and that it is processed by the general secretion (Sec) system. We further show that Ax21 is an outer membrane protein, secreted in association with outer membrane vesicles. Finally, we provide data showing that ax21 knockout strains do not overcome XA21-mediated immunity.

scholarly journals The peptidoglycan-associated protein NapA plays an important role in the envelope integrity and in the pathogenesis of the lyme disease spirochete

2021 ◽  
Vol 17 (5) ◽  
pp. e1009546
Author(s):  
Marisela M. Davis ◽  
Aaron M. Brock ◽  
Tanner G. DeHart ◽  
Brittany P. Boribong ◽  
Katherine Lee ◽  
...  
Keyword(s):  
Lyme Disease ◽  
Outer Membrane ◽  
Structural Integrity ◽  
Molecular Beacon ◽  
Cell Envelope ◽  
Protein A ◽  
Bacterial Pathogen ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles ◽  
Gram Negative

The bacterial pathogen responsible for causing Lyme disease, Borrelia burgdorferi, is an atypical Gram-negative spirochete that is transmitted to humans via the bite of an infected Ixodes tick. In diderms, peptidoglycan (PG) is sandwiched between the inner and outer membrane of the cell envelope. In many other Gram-negative bacteria, PG is bound by protein(s), which provide both structural integrity and continuity between envelope layers. Here, we present evidence of a peptidoglycan-associated protein (PAP) in B. burgdorferi. Using an unbiased proteomics approach, we identified Neutrophil Attracting Protein A (NapA) as a PAP. Interestingly, NapA is a Dps homologue, which typically functions to bind and protect cellular DNA from damage during times of stress. While B. burgdorferi NapA is known to be involved in the oxidative stress response, it lacks the critical residues necessary for DNA binding. Biochemical and cellular studies demonstrate that NapA is localized to the B. burgdorferi periplasm and is indeed a PAP. Cryo-electron microscopy indicates that mutant bacteria, unable to produce NapA, have structural abnormalities. Defects in cell-wall integrity impact growth rate and cause the napA mutant to be more susceptible to osmotic and PG-specific stresses. NapA-linked PG is secreted in outer membrane vesicles and augments IL-17 production, relative to PG alone. Using microfluidics, we demonstrate that NapA acts as a molecular beacon—exacerbating the pathogenic properties of B. burgdorferi PG. These studies further our understanding of the B. burgdorferi cell envelope, provide critical information that underlies its pathogenesis, and highlight how a highly conserved bacterial protein can evolve mechanistically, while maintaining biological function.

scholarly journals Proteins in the periplasmic space and outer membrane vesicles ofRhizobium etliCE3 grown in minimal medium are largely distinct and change with growth phase

2018 ◽  
Author(s):  
Hermenegildo Taboada ◽  
Niurka Meneses ◽  
Michael F. Dunn ◽  
Carmen Vargas-Lagunas ◽  
Natasha Buchs ◽  
...  
Keyword(s):  
Outer Membrane ◽  
Minimal Medium ◽  
Proteome Analysis ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles ◽  
Periplasmic Space ◽  
Rhizobium Etli ◽  
Multidrug Efflux ◽  
Secretion Systems ◽  
Shock Proteins

ABSTRACTRhizobium etliCE3 grown in succinate-ammonium minimal medium (MM) excreted outer membrane vesicles (OMVs) with diameters of 40 to 100 nm. Proteins from the OMVs and the periplasmic space were isolated from 6 and 24 h cultures and identified by proteome analysis. A total 770 proteins were identified: 73.8 and 21.3 % of these proteins occurred only in the periplasm and OMVs, respectively, and only 4.9 % were found in both locations. The majority of proteins found in either location were present only at 6 or 24 h: in the periplasm and OMVs, only 24 and 9 % of proteins, respectively, were present at both sampling times, indicating a time-dependent differential sorting of proteins into the two compartments. The OMVs contained proteins with physiologically varied roles, includingRhizobiumadhering proteins (Rap), polysaccharidases, polysaccharide export proteins, autoaggregation and adherence proteins, glycosyl transferases, peptidoglycan binding and cross-linking enzymes, potential cell wall modifying enzymes, porins, multidrug efflux RND family proteins, ABC transporter proteins, and heat shock proteins. As expected, proteins with known periplasmic localizations (phosphatases, phosphodiesterases, pyrophosphatases) were found only in the periplasm, along with numerous proteins involved in amino acid and carbohydrate metabolism and transport. Nearly one-quarter of the proteins present in the OMVs were also found in our previous analysis of theR. etlitotal exproteome of MM-grown cells, indicating that these nanoparticles are an important mechanism for protein excretion in this species.IMPORTANCEThe reduction of atmospheric nitrogen to ammonia by rhizobia symbiotically associated with legumes is of major importance in sustainable agricultural. Rhizobia excrete a variety of symbiotically important proteins using canonical secretion systems. In this work, we show thatRhizobium etligrown in culture also excretes proteins in membrane-enclosed structures called outer membrane vesicles (OMVs). This study reports OMV production by rhizobia. Proteins identified in the OMVs included Rhizobium adhering (Rap) and autoaggregation proteins, polysaccharidases, RTX toxins, porins and multidrug efflux proteins. Some of these proteins have important roles in theR. etli-common bean symbiosis, and their packaging into OMVs could deliver them to the environment in a concentrated yet diffusible form protected from degradation. The work described here provides a basis for future studies on the function of rhizobial OMVs in free life and symbiosis.

scholarly journals Characterization of Outer Membrane Vesicles fromBrucella melitensisand Protection Induced in Mice

2012 ◽  
Vol 2012 ◽  
pp. 1-13 ◽  
Author(s):  
Eric Daniel Avila-Calderón ◽  
Ahidé Lopez-Merino ◽  
Neeta Jain ◽  
Humberto Peralta ◽  
Edgar Oliver López-Villegas ◽  
...  
Keyword(s):  
Outer Membrane ◽  
Virulent Strain ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles ◽  
Saline Control ◽  
Control Group ◽  
Cell Mediated Immunity ◽  
Rough Strain ◽  
Smooth Strain

The outer membrane vesicles (OMVs) from smoothB. melitensis16 M and a derived rough mutant, VTRM1 strain, were purified and characterized with respect to protein content and induction of immune responses in mice. Proteomic analysis showed 29 proteins present in OMVs fromB. melitensis16 M; some of them are well-knownBrucellaimmunogens such as SOD, GroES, Omp31, Omp25, Omp19, bp26, and Omp16. OMVs from a rough VTRM1 induced significantly higher expression of IL-12, TNFα, and IFNγ genes in bone marrow dendritic cells than OMVs from smooth strain 16 M. Relative to saline control group, mice immunized intramuscularly with rough and smooth OMVs were protected from challenge with virulent strainB. melitensis16 M just as well as the group immunized with live strainB. melitensisRev1 (P<0.005). Additionally, the levels of serum IgG2a increased in mice vaccinated with OMVs from rough strain VTRM1 consistent with the induction of cell-mediated immunity.

scholarly journals Characterization and Vaccine Potential of Outer Membrane Vesicles Produced by Haemophilus parasuis

PLoS ONE ◽  
2016 ◽  
Vol 11 (3) ◽  
pp. e0149132 ◽  
Author(s):  
William D. McCaig ◽  
Crystal L. Loving ◽  
Holly R. Hughes ◽  
Susan L. Brockmeier
Keyword(s):  
Outer Membrane ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles ◽  
Haemophilus Parasuis ◽  
Vaccine Potential

scholarly journals The Xanthomonas Ax21 protein is processed by the general secretory system and is secreted in association with outer membrane vesicles

2013 ◽  
Author(s):  
Ofir Bahar ◽  
Rory N Pruitt ◽  
Dee Dee Luu ◽  
Benjamin Schwessinger ◽  
Randy Ruan ◽  
...  
Keyword(s):  
Outer Membrane ◽  
Bacterial Pathogen ◽  
Membrane Vesicles ◽  
Secretion System ◽  
Outer Membrane Vesicles ◽  
Type I ◽  
Broad Spectrum Resistance ◽  
Sec System ◽  
Multiple Strains ◽  
Secretory System

Pattern recognition receptors (PRRs) play an important role in detecting invading pathogens and mounting a robust defense response to restrict infection. In rice, one of the best characterized PRRs is XA21, a leucine rich repeat receptor-like kinase that confers broad-spectrum resistance to multiple strains of the bacterial pathogen Xanthomonas oryzae pv. oryzae (Xoo). In 2009 we reported that an Xoo protein, called Ax21, was secreted by a type I-secretion system and that it served to a ctivate X A 21 -mediated immunity. This report has recently been retracted. Here we present data that corrects our previous model. We first show that Ax21 secretion does not depend on the predicted type I secretion system and that it is processed by the general secretion (Sec) system. We further show that Ax21 is an outer membrane protein, secreted in association with outer membrane vesicles. Finally, we provide data showing that ax21 knockout strains do not overcome XA21-mediated immunity.

scholarly journals Deletion of alpB Gene Influences Outer Membrane Vesicles Biogenesis of Lysobacter sp. XL1

2021 ◽  
Vol 12 ◽  
Author(s):  
Irina V. Kudryakova ◽  
Alexey S. Afoshin ◽  
Tanya V. Ivashina ◽  
Natalia E. Suzina ◽  
Elena A. Leontyevskaya ◽  
...  
Keyword(s):  
Outer Membrane ◽  
Wild Type Strain ◽  
Membrane Vesicles ◽  
Outer Membrane Vesicles ◽  
Lower Amount ◽  
Extracellular Milieu ◽  
Allelic Replacement ◽  
Vesicle Biogenesis ◽  
Ultrastructural Characteristics ◽  
Genetic Deletion

Outer membrane vesicles (OMVs) produced by Gram-negative bacteria constitute important factors in defining interactions with the extracellular milieu. Lysobacter sp. XL1 produces OMVs capable of lysing microbial cells due to the presence in their cargo of bacteriolytic protease L5 (AlpB). Although protein L5 has been functionally and biochemically characterized (including aspects of its packing into OMVs), its role in vesicle biogenesis through genetic deletion of alpB had not been studied previously. Here, we have successfully deleted alpB by allelic replacement and show that the alpB deletion mutant produces a significantly lower amount of OMVs that lack bacteriolytic activity and display altered ultrastructural characteristics in relation to the OMVs produced by the wild-type strain. These results confirm that, as previously proposed, protein L5 participates in OMV production through a mechanism that is not yet fully understood.

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