Investigation of Protein Fatty Acylation in Virus-Host Interaction

Fatty acylation has a critical impact on antiviral innate immunity. Fatty acylation of virus proteins affects virus virulence and invasion. In contrast, fatty acylation of host cell proteins regulates virus recognition and host-virus interactions, thus affecting antiviral immune responses. Therefore, an emerging need exists to investigate and understand the mechanism and function of protein fatty acylation in regulating antiviral innate immunity. Alkyne probe metabolic labeling and click chemistry assays with biotin-conjugated or fluorophore-conjugated azide provide a simple way to determine the potential fatty-acylated proteins in antiviral innate pathways. Here, we demonstrate comprehensive approaches to determine protein fatty acylation in virus-infected macrophages or protein-overexpressing HEK293T cells in the context of antiviral innate immunity.