Quantitative Proteomics by Amino Acid Labeling in Foot-and-Mouth Disease Virus (FMDV)-Infected Cells

Abstract

Foot-and-mouth disease virus (FMDV) is an important disease agent that can be difficult to effectively eradicate from herds. Because it is an obligate intracellular parasite, the virus has multiple effects on the host cell during infection. Here, a high-throughput quantitative proteomic approach was used to develop an unbiased holistic overview of the protein changes in IBRS-2 cells infected with FMDV. Stable isotope labeling with amino acids in cell culture (SILAC) combined with LC-MS/MS was performed to identify and quantify 1260 cellular and 2 viral proteins after 6 h of infection of IBRS-2 cells with FMDV. Of these identified and measured cellular protein pairs, 77 were significantly up-regulated, and 50 were significantly down-regulated based on significance B ≤ 0.05. The differentially altered proteins included a number of proteins involved in endolysosomal proteases system, cell cycle, cellular growth and proliferation, and immune cell trafficking. Selected data were validated by Western blot. Ingenuity Pathway Analysis revealed that proteins that changed in response to infection could be assigned to defined canonical pathways and functional groupings, such as integrin signaling. The obtained data might not only improve the understanding of the dynamics of FMDV and host interaction but may also help elucidate the pathogenic mechanism of FMDV infection.

The pig as an animal model for human pathologies: A proteomics perspective

“…Haptoglobin and apolipoprotein A‐IV precursor were detectable in serum after infection, the former having been previously identified in FMDV‐infected calves . Intra‐cellular responses of IBRS‐2 cells to the FMDV have been demonstrated by SILAC coupled with LC‐MS/MS , revealing 77 proteins that were up‐regulated and 50 that were down‐regulated in a group of 1260 cellular proteins in total. The proteins altered by infection were involved in endolysosomal protease action, cell cycle, cellular growth and immune cell trafficking among other activities and provided a novel insight into the cellular interactions following FMDV infection.…”

Three-dimensional structure of foot-and-mouth disease virus and its biological functions

“…The development of high-throughput screening technologies to understand FMDV-host interactions has considerably enriched our knowledge of viral infections [88,149]. Structural methods (X-ray crystallography, nuclear magnetic resonance, and electron microscopy) combined with various '-omics' approaches (transcriptomics, proteomics, etc.)…”

An extracellular laccase with potent dye decolorizing ability from white rot fungus Trametes sp. LAC-01

“…The data were acquired using Xcalibur software (Thermo Electron). Sequence homologues were searched using the BLAST/NCBI database [25,26].…”