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In silico identification of potential drug targets in swine pathogen Haemophilus parasuis
Zhuofei Xu,Lifang Ma,Huanchun Chen,Rui Zhou 한국유전학회 2012 Genes & Genomics Vol.34 No.2
Gram-negative bacterium Haemophilus parasuis has recently become one of the most important etiological agents causing serious systemic disease (Glässer’s disease) in pigs. Antibiotic therapy has played a crucial role in the treatment of this disease. Antibiotic resistance observed from the clinical isolates of this pathogen urges us to discover novel drug targets for antimicrobial agents. In this study, we used a combined strategy including exploration of the gene essentiality and comparison of metabolic pathways to infer drug targets of H. parasuis. We identified 931 gene products essential for bacterial growth according to the DEG database. One hundred and ninety-nine enzyme-coding genes were found in the genome of H. parasuis but were absent in that of the swine host. Lastly,we determined 117 enzymes exhibiting essentiality and specificity to H. parasuis as a candidate set of drug targets. Comparison of metabolic pathways between the pathogen and host showed that 25 targeting enzymes belonged to nine unique pathways of the pathogen. The profile of promising targets identified in our study will provide a useful basis for developing more effective antibiotics against the severe swine disease caused by H. parasuis.
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Xiabing Chen,Zhuofei Xu,Lu Li,Huanchun Chen,Rui Zhou 한국미생물학회 2012 The journal of microbiology Vol.50 No.6
Actinobacillus pleuropneumoniae is an important swine respiratory pathogen causing great economic losses worldwide. Identification of conserved surface antigenic proteins is helpful for developing effective vaccines. In this study, a genome-wide strategy combined with bioinformatic and experimental approaches, was applied to discover and characterize surface-associated immunogenic proteins of A. pleuropneumoniae. Thirty nine genes encoding outer membrane proteins (OMPs) and lipoproteins were identified by comparative genomics and gene expression profiling as beinghighly conserved and stably transcribed in the different serotypes of A. pleuropneumoniae reference strains. Twelve of these conserved proteins were successfully expressed in Escherichia coli and their immunogenicity was estimated by homologous challenge in the mouse model, and then three of these proteins (APJL_0126, HbpA and OmpW) were further tested in the natural host (swine) by homologous and heterologous challenges. The results showed that these proteins could induce high titers of antibodies, but vaccination with each protein individually elicited low protective immunity against A. pleuropneumoniae. This study gives novel insights into immunogenicity of the conserved OMPs and lipoproteins of A. pleuropneumoniae. Although none of the surface proteins characterized in this study could individually induce effective protective immunity against A. pleuropneumoniae, they are potential candidates for subunit vaccines in combination with Apx toxins.
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Lu Li,Jiawen Zhu,Kui Yang,Zhuofei Xu,Ziduo Liu,Rui Zhou 한국미생물학회 2014 The journal of microbiology Vol.52 No.6
Actinobacillus pleuropneumoniae is an important porcinerespiratory pathogen causing great economic losses in thepig industry worldwide. Oxygen deprivation is a stress thatA. pleuropneumoniae will encounter during both early infectionand the later, persistent stage. To understand modulationof A. pleuropneumoniae gene expression in responseto the stress caused by anaerobic conditions, gene expressionprofiles under anaerobic and aerobic conditions werecompared in this study. The microarray results showed that631 genes (27.7% of the total ORFs) were differentially expressedin anaerobic conditions. Many genes encoding proteinsinvolved in glycolysis, carbon source uptake systems,pyruvate metabolism, fermentation and the electron respirationtransport chain were up-regulated. These changes ledto an increased amount of pyruvate, lactate, ethanol and acetatein the bacterial cells as confirmed by metabolite detection. Genes encoding proteins involved in cell surface structures,especially biofilm formation, peptidoglycan biosynthesisand lipopolysaccharide biosynthesis were up-regulatedas well. Biofilm formation was significantly enhancedunder anaerobic conditions. These results indicate that inductionof central metabolism is important for basic survivalof A. pleuropneumoniae after a shift to an anaerobic environment. Enhanced biofilm formation may contribute tothe persistence of this pathogen in the damaged anaerobichost tissue and also in the early colonization stage. Thesediscoveries give new insights into adaptation mechanismsof A. pleuropneumoniae in response to environmental stress.
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Siqi Li,Zhipeng Su,Chengjun Zhang,Zhuofei Xu,Xiaoping Chang,Jiawen Zhu,Ran Xiao,Lu Li,Rui Zhou 한국유전학회 2018 Genes & Genomics Vol.40 No.8
Porcine pleuropneumonia caused by Actinobacillus pleuropneumoniae has led to severe economic losses in the pig industry worldwide. A. pleuropneumoniae displays various levels of antimicrobial resistance, leading to the dire need to identify new drug targets. Protein–protein interaction (PPI) network can aid the identification of drug targets by discovering essential proteins during the life of bacteria. The aim of this study is to identify drug target candidates of A. pleuropneumoniae from essential proteins in PPI network. The homologous protein mapping method (HPM) was utilized to construct A. pleuropneumoniae PPI network. Afterwards, the subnetwork centered with H-NS was selected to verify the PPI network using bacterial two-hybrid assays. Drug target candidates were identified from the hub proteins by analyzing the topology of the network using interaction degree and homologous comparison with the pig proteome. An A. pleuropneumoniae PPI network containing 2737 non-redundant interaction pairs among 533 proteins was constructed. These proteins were distributed in 21 COG functional categories and 28 KEGG metabolic pathways. The A. pleuropneumoniae PPI network was scale free and the similar topological tendencies were found when compared with other bacteria PPI network. Furthermore, 56.3% of the H-NS subnetwork interactions were validated. 57 highly connected proteins (hub proteins) were identified from the A. pleuropneumoniae PPI network. Finally, 9 potential drug targets were identified from the hub proteins, with no homologs in swine. This study provides drug target candidates, which are promising for further investigations to explore lead compounds against A. pleuropneumoniae.
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