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Updated: Jul 15, 2026
Characterization at the Molecular Level using Robust Biochemical Approaches of a New Kinase Protein
Published on: June 30, 2019
IP-LC-MS/MS-Based Identification of Candidate TPL2 (MAP3K8)-Associated Proteins in Porcine PK-15 Cells
Dajun Zhang1, Dengshuai Zhao1, Jiajun Lin1
1School of Animal Science and Technology, Foshan University, Foshan, China.
Abstract:
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.
Insights
Researchers identified 666 proteins interacting with Tumor Progression Locus 2 (TPL2) in swine cells using IP-LC-MS/MS. This dataset aids understanding TPL2
Area of Science:
- Immunology
- Molecular Biology
- Proteomics
Background:
- Tumor Progression Locus 2 (TPL2) is a kinase crucial for MAPK and NF-κB signaling pathways, impacting antiviral immunity.
- While TPL2's role in mammals is well-documented, its protein interactions in swine cells are largely uncharacterized.
Purpose of the Study:
- To identify and characterize proteins associated with TPL2 in porcine kidney epithelial (PK-15) cells.
- To establish a foundational dataset for investigating TPL2's function in swine host-pathogen interactions.
Main Methods:
- Overexpression of FLAG-tagged TPL2 in PK-15 cells.
- Immunoprecipitation (IP) to enrich TPL2-associated proteins.
- Liquid chromatography-tandem mass spectrometry (LC-MS/MS) for protein identification.
- Bioinformatic analyses including GO, KEGG pathway, and PPI network analysis.
Main Results:
- A dataset of 666 candidate TPL2-associated proteins was generated after removing background noise from negative controls.
- Bioinformatic analyses revealed enrichment in RNA binding, protein modification, ubiquitin-proteasome pathways, spliceosome, and immune signaling.
- Identified proteins suggest TPL2's involvement in diverse cellular processes beyond its known kinase activity.
Conclusions:
- This study presents the first comprehensive dataset of TPL2-interacting proteins in porcine cells.
- The findings provide a valuable resource for future research into TPL2-mediated signaling and its role in swine antiviral immunity and virus-host interactions.
