Demonstration of the third antigenically distinct outer membrane lipoprotein (OmlA) in Actinobacillus pleuropneumoniae serotype 7

Ito, Hiroya

doi:10.1016/s0378-1097(98)00407-8

Cited by 3 publications

(3 citation statements)

References 8 publications

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“…OmlA is expressed by isolates of all A. pleuropneumoniae serotypes, but sequence variation between alleles results in three antigenically distinct forms. OmlA 1 is present in serotypes 1, 2, 8, 9, 11, and 12 (18); OmlA 5 in serotypes 5a, 5b, and 10 (11, 23); and OmlA 7 in serotypes 3, 4, 6, and 7 (22). The anti-OmlA antiserum used in this study was raised against OmlA 1 and does not recognize the OmlA 5 and OmlA 7 variants in immunoblots (18), preventing us from determining conclusively whether AasP-mediated OmlA processing occurs in strains expressing these variants.…”

Section: Discussionmentioning

confidence: 99%

Functional Characterization of AasP, a Maturation Protease Autotransporter Protein ofActinobacillus pleuropneumoniae

et al. 2008

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Actinobacillus pleuropneumoniae is the etiological agent of porcine pleuropneumonia, a highly contagious respiratory infection in pigs. AasP, a putative subtilisin-like serine protease autotransporter, has recently been identified in A. pleuropneumoniae. We hypothesized that, similarly to other autotransporters of this type, AasP may undergo autocatalytic cleavage resulting in release of the passenger domain of the protein. Furthermore, AasP may be responsible for cleavage of other A. pleuropneumoniae outer membrane proteins. To address these hypotheses, the aasP gene was cloned and the expressed recombinant AasP protein used to raise monospecific rabbit antiserum. Immunoblot analysis of whole-cell lysates and secreted proteins demonstrated that AasP does not undergo proteolytic cleavage. Immunoblot analysis also confirmed that AasP is universally expressed by A. pleuropneumoniae. Confirmation of the maturation protease function of AasP was obtained through phenotypic analysis of an A. pleuropneumoniae aasP deletion mutant and by functional complementation. Comparison of the secreted proteins of the wild type, an aasP mutant derivative, and an aasP mutant complemented in trans led to the identification of OmlA protein fragments that were present only in the secreted-protein preparations of the wild-type and complemented strains, indicating that AasP is involved in modification of OmlA. This is the first demonstration of a function for any autotransporter protein in Actinobacillus pleuropneumoniae.

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Section: Discussionmentioning

confidence: 99%

Functional Characterization of AasP, a Maturation Protease Autotransporter Protein ofActinobacillus pleuropneumoniae

et al. 2008

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“…A new serotyping method using Western blot analysis with specific antisera against three antigenically distinct OmlA proteins has been developed 16,17 . The Western blot analysis using specific antisera against OmlA 5a(NG-8) (Fig.…”

Section: Development Of Typing Methods Based On the Antigenic Diversitmentioning

confidence: 99%

Development of Typing Methods of Actinobacillus pleuropneumoniae Based on the Antigenic and Genetic Diversity of the Protective Outer Membrane Lipoprotein

Ito

2008

JARQ

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Actinobacillus pleuropneumoniae is a causative agent of porcine pleuropneumonia that is one of the important bacterial diseases of the respiratory tract of pigs. The disease occurs worldwide and results in serious economic losses to the pig-rearing industry. In this review, new typing methods based on the antigenic and genetic diversity of a protective outer membrane lipoprotein (OmlA), that have been developed for control and epidemiological study of the disease, are described. First, the gene for the OmlA of an A. pleuropneumoniae field isolate (serotype 5a) has been cloned and characterized. The lipoprotein has been shown to be a protective antigen of the organism. Southern blot and the nucleotide sequence analysis of the gene for the lipoprotein have revealed the presence and diversity of the gene in A. pleuropneumoniae. Secondly, based on the antigenic diversity of OmlA, Western blot analysis with specific antisera against three antigenically distinct OmlAs has been developed. The antisera can differentiate A. pleuropneumoniae into three groups. Finally, based on the genetic diversity of OmlA, a PCR-Restriction Fragment Length Polymorphism (RFLP) typing method has been developed. This method can divide A. pleuropneumoniae into five groups. The grouping results by the Western blot analysis were correlated with that by the PCR-RFLP typing method. In addition, the PCR-RFLP typing method can be performed in almost all veterinary diagnostic laboratories since the method does not require specific antisera and expensive reagents and apparatus. Therefore, the PCR-RFLP typing method can be substituted for other OmlA typing and seems to be the most convenient and practical OmlA typing method at present. The new typing methods developed in this study should be used as tools which are effective and practical for control of porcine pleuropneumonia caused by A. pleuropneumoniae.

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“…For A. pleuropneumoniae, several putative virulence-associated factors such as the RTX toxins ApxI, ApxII, and ApxIII (2,6,12,19,20,24,33,37), the transferrin binding proteins TbpA and TbpB (13,15,25,41), a superoxide dismutase (SodA [23]), a urease (4), an outer membrane lipoprotein (OmlA [5,14,17,18]), and capsular antigens (16,40) have been characterized. The RTX toxins are distributed in a serotype-specific manner with each serotype containing no more than two of the Apx toxins (12,20), and the TbpB and OmlA proteins occur in three antigenically distinct isoforms (13,17).…”

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First Chromosomal Restriction Map ofActinobacillus pleuropneumoniaeand Localization of Putative Virulence-Associated Genes

et al. 1999

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Combined physical and genetic maps of the genomes ofActinobacillus pleuropneumoniae AP76 (serotype 7 clinical isolate) and of A. pleuropneumoniae ATCC 27088 (serotype 1 reference strain) were constructed by using the restriction endonucleases ApaI, AscI, NotI, andSalI. The chromosome sizes as determined by the addition of estimated fragment sizes were 2.4 Mbp, and both maps had a resolution of approximately 100 kbp. The linkages between the ApaI,AscI, NotI, and SalI fragments and their relative positions were determined by (i) fragment excision and redigestion and (ii) partial digests of defined fragments and Southern blot using end-standing probes. The single SalI site within the chromosome of strain A. pleuropneumoniae AP76 was defined as position 1 of the map; for the map of A. pleuropneumoniae ATCC 27088, the corresponding SalI site was chosen. Putative virulence-associated genes (apx,omlA, sodA, tbpBA,ureC, and a repeat element) and housekeeping genes (glyA, metJ, recA, andrhoAP) were positioned on the physical maps and located on the ApaI and NotI fragments of A. pleuropneumoniae serotype reference strains.

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Demonstration of the third antigenically distinct outer membrane lipoprotein (OmlA) in Actinobacillus pleuropneumoniae serotype 7

Cited by 3 publications

References 8 publications

Functional Characterization of AasP, a Maturation Protease Autotransporter Protein ofActinobacillus pleuropneumoniae

Functional Characterization of AasP, a Maturation Protease Autotransporter Protein ofActinobacillus pleuropneumoniae

Development of Typing Methods of Actinobacillus pleuropneumoniae Based on the Antigenic and Genetic Diversity of the Protective Outer Membrane Lipoprotein

First Chromosomal Restriction Map ofActinobacillus pleuropneumoniaeand Localization of Putative Virulence-Associated Genes

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