Actinobacillus pleuropneumoniae metalloprotease: cloning and in vivo expression

González, Octavio Garcı́a

doi:10.1016/j.femsle.2004.03.012

Cited by 5 publications

(5 citation statements)

References 17 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…It was characterized on the basis of a zinc binding motif (aa 294-303) found in metalloproteases. Expression of this protease was observed in lung tissue of pigs that had died from porcine pleuropneumonia [45] .…”

Section: Resultsmentioning

confidence: 99%

Genome Biology of Actinobacillus pleuropneumoniae JL03, an Isolate of Serotype 3 Prevalent in China

Zhao

Zhou

et al. 2008

PLoS ONE

View full text Add to dashboard Cite

Actinobacillus pleuropneumoniae is the etiologic agent of porcine contagious pleuropneumonia, a cause of considerable world wide economic losses in the swine industry. We sequenced the complete genome of A. pleuropneumoniae, JL03, an isolate of serotype 3 prevalent in China. Its genome is a single chromosome of 2,242,062 base pairs containing 2,097 predicted protein-coding sequences, six ribosomal rRNA operons, and 63 tRNA genes. Preliminary analysis of the genomic sequence and the functions of the encoded proteins not only confirmed the present physiological and pathological knowledge but also offered new insights into the metabolic and virulence characteristics of this important pathogen. We identified a full spectrum of genes related to its characteristic chemoheterotrophic catabolism of fermentation and respiration with an incomplete TCA system for anabolism. In addition to confirming the lack of ApxI toxin, identification of a nonsense mutation in apxIVA and a 5′-proximal truncation of the flp operon deleting both its promoter and the flp1flp2tadV genes have provided convincing scenarios for the low virulence property of JL03. Comparative genomic analysis using the available sequences of other serotypes, probable strain (serotype)-specific genomic islands related to capsular polysaccharides and lipopolysaccharide O-antigen biosyntheses were identified in JL03, which provides a foundation for future research into the mechanisms of serotypic diversity of A. pleuropneumoniae.

show abstract

Section: Resultsmentioning

confidence: 99%

Genome Biology of Actinobacillus pleuropneumoniae JL03, an Isolate of Serotype 3 Prevalent in China

Zhao

Zhou

et al. 2008

PLoS ONE

View full text Add to dashboard Cite

show abstract

“…A. pleuropneumoniae represents no exception, and for instance the inactivation of the hgbA gene has been linked to reduced virulence in vivo [ 54 ]. PepN is another interesting candidate immunogen in our list, a protease able to degrade porcine IgA and IgG [ 55 ]. It is intuitive to understand how targeting antibody degradation could be beneficial in reducing A. pleuropneumoniae resistance to the host immune system.…”

Section: Discussionmentioning

confidence: 99%

Identification and characterization of serovar-independent immunogens in Actinobacillus pleuropneumoniae

Antenucci

Fougeroux

Bossé

et al. 2017

Vet Res

View full text Add to dashboard Cite

Despite numerous actions to prevent disease, Actinobacillus pleuropneumoniae (A. pleuropneumoniae) remains a major cause of porcine pleuropneumonia, resulting in economic losses to the swine industry worldwide. In this paper, we describe the utilization of a reverse vaccinology approach for the selection and in vitro testing of serovar-independent A. pleuropneumoniae immunogens. Potential immunogens were identified in the complete genomes of three A. pleuropneumoniae strains belonging to different serovars using the following parameters: predicted outer-membrane subcellular localization; ≤ 1 trans-membrane helices; presence of a signal peptide in the protein sequence; presence in all known A. pleuropneumoniae genomes; homology with other well characterized factors with relevant data regarding immunogenicity/protective potential. Using this approach, we selected the proteins ApfA and VacJ to be expressed and further characterized, both in silico and in vitro. Additionally, we analysed outer membrane vesicles (OMVs) of A. pleuropneumoniae MIDG2331 as potential immunogens, and compared deletions in degS and nlpI for increasing yields of OMVs compared to the parental strain. Our results indicated that ApfA and VacJ are highly conserved proteins, naturally expressed during infection by all A. pleuropneumoniae serovars tested. Furthermore, OMVs, ApfA and VacJ were shown to possess a high immunogenic potential in vitro. These findings favour the immunogen selection protocol used, and suggest that OMVs, along with ApfA and VacJ, could represent effective immunogens for the prevention of A. pleuropneumoniae infections in a serovar-independent manner. This hypothesis is nonetheless predictive in nature, and in vivo testing in a relevant animal model will be necessary to verify its validity.Electronic supplementary materialThe online version of this article (10.1186/s13567-017-0479-5) contains supplementary material, which is available to authorized users.

show abstract

“…[ 31 ]. The membranes were blocked with skim milk for 2 h at 22 °C, and the samples were incubated overnight with a rabbit anti- A. pleuropneumoniae 100 kDa protease (1 : 2000) antiserum at 4 °C [ 32 ]. The samples were washed three times with PBS-Tween (0.05 %) and incubated with a secondary anti-rabbit-HRP antibody (1 : 3000) (Sigma) for 1 h at 22 °C.…”

Section: Methodsmentioning

confidence: 99%

Bovine apo-lactoferrin affects the secretion of proteases in Mannheimia haemolytica A2

Ramírez-Rico

Martínez-Castillo

Avalos-Gómez

et al. 2021

Access Microbiology

View full text Add to dashboard Cite

Mannheimia haemolytica serotype A2 is the main bacterial causative agent of ovine mannheimiosis, a disease that leads to substantial economic losses for livestock farmers. Several virulence factors allow M. haemolytica to colonize the lungs and establish infection. Virulence factors can be directly secreted into the environment by bacteria but are also released through outer membrane vesicles (OMVs). In addition, due to the abuse of antibiotics in the treatment of this disease, multidrug-resistant bacterial strains of M. haemolytica have emerged. One therapeutic alternative to antibiotics or an adjuvant to be used in combination with antibiotics could be lactoferrin (Lf), a multifunctional cationic glycoprotein of the mammalian innate immune system to which no bacterial resistance has been reported. The aim of this work was to determine the effect of bovine iron-free Lf (apo-BLf) on the production and secretion of proteases into culture supernatant (CS) and on their release in OMVs. Zymography assays showed that addition of sub-MIC concentrations of apo-BLf to M. haemolytica cultures inhibited protease secretion without affecting culture growth. Biochemical characterization revealed that these proteases were mainly cysteine- and metalloproteases. The secretion of a 100 kDa metalloprotease was inhibited by sub-MIC concentrations of apo-BLf since this protease was present in the cytoplasm and OMVs but not in CS proteins, as corroborated by Western blotting. On the other hand, proteases produced by M. haemolytica caused cleavage of apo-BLf. However, when Lf is cleaved, peptides known as lactoferricins, which are more bactericidal than natural Lf, can be produced. M. haemolytica A2 protease-mediated degradation of host tissue proteins could be an important virulence factor during the infectious process of pneumonia in ovines. The mechanism of M. haemolytica protease secretion could be inhibited by treatment with apo-BLf in animals.

show abstract

Actinobacillus pleuropneumoniae metalloprotease: cloning and in vivo expression

Cited by 5 publications

References 17 publications

Genome Biology of Actinobacillus pleuropneumoniae JL03, an Isolate of Serotype 3 Prevalent in China

Genome Biology of Actinobacillus pleuropneumoniae JL03, an Isolate of Serotype 3 Prevalent in China

Identification and characterization of serovar-independent immunogens in Actinobacillus pleuropneumoniae

Bovine apo-lactoferrin affects the secretion of proteases in Mannheimia haemolytica A2

Contact Info

Product

Resources

About