Quantitative Proteomic Analysis of Global Protein Acetylation in PRRSV‐Infected Pulmonary Alveolar Macrophages

Abstract

Porcine reproductive and respiratory syndrome (PRRS), caused by PRRS virus (PRRSV), is a serious viral disease affecting global swine industry. Due to the lack of effective vaccines, new antiviral strategies to compensate for the inefficacy of available vaccines are urgently required. Lysine acetylation, as an important post‐translational modification during infection, plays a key regulatory role in host antiviral responses. In this study, the global acetylome is profiled using acetylation specific antibody‐based enrichment and tandem mass tag label high‐affinity purification liquid chromatography‐mass spectrometry in PRRSV‐infected pulmonary alveolar macrophages (PAMs). As a result, 3731 lysine acetylation sites on 1421 cellular proteins are identified. Bioinformatics analysis of the different acetylated proteins revealed their involvement in various biological processes, including the host immune response and energy metabolism. These findings will contribute to the understanding of PRRSV pathogenesis and identify new cellular targets for anti‐PPRSV therapeutics.

Mechanism of interaction between virus and host is inferred from the changes of gene expression in macrophages infected with African swine fever virus CN/GS/2018 strain

“…PAMs are an immune cells with strong chemotaxis, can secrete a variety of chemokines, and attract and activates immune cells to enhance the immune and inflammatory response [ 23 ]. Therefore, combined with the data of RNA-seq, the transcriptional level of CCL4 (C-C motif chemokine 4) , CCL5, CXCL8, and CXCL10 in PAMs infected with ASFV was detected by qPCR at different time points.…”

A proteomic view of cellular responses of macrophages to copper when added as ion or as copper-polyacrylate complex

“…It may be argued that enzymes may be regulated differently from other cellular proteins. However, the scope of proteins identified in phosphoproteome [62] or acetylome [63] studies extends far beyond enzymes, so that assuming that enzymes are not regulated by different mechanisms than other proteins appears reasonable. This suggests that the statistical values obtained when comparing proteomic to activity results for enzymes may be extended globally to the complete proteomic experiment.…”

“…The instrument was used with a MicroMist U-Series glass concentric nebulizer, a quartz spray chamber cooled at 3°C, a Qnova quartz torch, a nickel sample cone, and a nickel skimmer cone equipped with a high-sensitivity insert. Elements were analyzed using kinetic energy discrimination mode with helium as the collision cell gas (for 63 Cu and 65 Cu). Concentrations were determined using standard curves and corrected using an internal standard solution of 103Rh added online.…”

“…Data integration was done using the Qtegra software (version 2.8.2944.115). The background equivalent concentration (BEC) on pure standards was 0.02 ng/ml for 63 Cu and 65 Cu.…”

Proteomic Investigation Reveals Eukaryotic Translation Initiation Factor 5A Involvement in Porcine Reproductive and Respiratory Syndrome Virus Infection in vitro

“…For example, Fang et al . ( 16 ) used an acetylation-based antibody enrichment technique and a tandem mass tag label high-affinity purification liquid chromatography-mass spectrometry (LC-MS/MS) method to study acetylome regulation of antiviral activities in PRRSV-infected PAMs. A few years earlier, Zhang et al ( 17 ) had generated a broad-spectrum ubiquitination modification map of PRRSV-infected PAMs using ubiquitination antibody enrichment in combination with MS technology.…”