IP-LC-MS/MS-Based Identification of Candidate TPL2 (MAP3K8)-Associated Proteins in Porcine PK-15 Cells

Tumor progression locus 2 (TPL2, also known as MAP-3K8/COT) is a kinase that regulates MAPK and NF-κB signaling and contributes to antiviral immunity. While mammalian TPL2 has been widely studied, the TPL2-associated protein profile in swine cells remains largely unexplored. In this study, we generated an IP-LC-MS/MS-based candidate dataset of TPL2-associated proteins in porcine kidney epithelial (PK-15) cells. FLAG-tagged TPL2 was overexpressed in PK-15 cells and enriched by immunoprecipitation, followed by liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. After subtraction of proteins detected in the empty-vector negative IP control, a total of 666 candidate TPL2-associated proteins were identified. Bioinformatic analyses, including Gene Ontology (GO), Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment, and protein-protein interaction (PPI) network analysis, indicated that these candidates were mainly associated with RNA binding, protein modification, ubiquitin-proteasome processes, spliceosome-related pathways, and immune-associated signaling. Collectively, our results provide an IP-LC-MS/MS-based candidate resource of TPL2-associated proteins in porcine PK-15 cells, offering a useful basis for future studies on TPL2-mediated host signaling and virus-host interactions.