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Differential Phosphoproteome of Porcine Alveolar Macrophage Associated with Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus Infection

Author: WuJiang
Tutor: ZhangDeLi
School: Northwest University of Science and Technology
Course: Animal Biotechnology
Keywords: HuN4 strains of HP-PRRSV Porcine alveolar macrophages Differences in phosphorylation of proteins
CLC: S858.28
Type: Master's thesis
Year: 2011
Downloads: 79
Quote: 1
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Abstract


Porcine reproductive and respiratory syndrome (Porcine reproductive and respiratory syndrome, PRRS) is a porcine reproductive and respiratory syndrome virus (Porcine reproductive and respiratory syndrome virus, PRRSV) due to the global pig important infectious diseases. PRRSV has antigenic variability, addicted to macrophages, and antibody-dependent enhancement the role (ADE) and persistent infection characterized and compared with the classic PRRSV HP-of PRRSV of virulence significantly enhanced its virulence enhancement molecules mechanism is as yet no explanation. Protein is exercised cell life activities of the basic molecular phosphorylation is an important protein chemical modification, to play an important role in the play or a change in protein function. Virus infect host cells as an external signal, it will affect the cell's normal life activities through a series of cell signaling, protein phosphorylation of the signaling pathways in the broadest, most important post-translational modification. The main target cells in vivo, PRRSV in porcine alveolar macrophages (Porcine alveolar macrophage, PAM), HP-PRRSV infection of the host cell after difference analysis of phosphorylated proteins, contribute to the further study of HP-PRRSV high Rev. The molecular mechanism of the disease force data provide a valuable reference. This study selected three 30-day-old PRRSV-negative and antibody-negative healthy piglets, pig used to collect the PAM. Collected cells were divided into a control group and infection group, to infection cells collected 2 h after HuN4 strains of HP-PRRSV infection. Phosphorylated protein enrichment the kit (Qigen) successful phosphorylated proteins in the two cell groups, analysis by liquid chromatography tandem mass spectrometry (LC-MS/MS) rev with BioworksBrowser. 3.3 software in a non-redundant MS / MS International Protein Index (IPI) pig spectral data database retrieval to the control group of 420 of the 306 proteins in the infected group protein. Application DAVID Bioinformatics Resources 6.7 (NIAID / NIH) database search identified 113 proteins in the control group, functional annotation, including unique protein 27; infection group 145 protein functional annotation, including unique protein 59 a. At the same time, check the library set STY dynamically modified peptides 79.9799 retrieve the control group 58 phosphorylation sites belonging to 56 protein; 48 phosphorylation sites of peptides, belonging to 46 protein infected group. Application DAVID Bioinformatics Resources 6.7 (NIAID / NIH) the database Gene_Ontology analysis, the control group, according to the function of proteins and functional annotation of proteins in the infected group is divided into the immune response, stress response, signal transduction, protein metabolism, nucleic acid metabolism, metabolism, cell activity components of the cytoskeleton and organelles nine groups. Kyoto Encyclopedia of Genes and Genomes (Kyoto Encyclopedia of Genes and Genomes, KEGG) database analysis of the control group and the infected group two groups differentially expressed proteins involved in intracellular signaling pathways functional annotation of proteins involved in the control group to 6 signaling pathway in the infected group functional annotation of proteins involved in 5 signaling pathway. Phosphorylated proteins in the control group, we found the six specific membrane protein or a receptor protein: CD163, CD45, CD9, TLR5, Pglyrp2/pPGRP-LB LOX-1/OLR1; phosphorylated proteins in the infected group, we found that involved in the actin cytoskeleton regulation, there are significant differences in antigen processing and presentation and proteasome pathway protein. The exact role of these differences phosphorylated proteins play in the HP-PRRSV infection of PAM, yet to be confirmed by further studies.

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CLC: > Agricultural Sciences > Livestock, animal medicine,hunting,silkworm,bee > Animal Medicine ( Veterinary Medicine) > Livestock, poultry, wildlife diseases > Livestock > Pig
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