Identification of Toll-like receptor 4 interacting partner through immunoprecipitation-based chemical cross-linking proteomics

Toll-like receptor 4 (TLR-4) is a transmembrane protein. Its activation leads to NF-kB signaling pathway and proinflammatory cytokines’ production that are responsible for activating innate immunity system. Here, we design a co-immunoprecipitation based cross-linking proteomics approach to reveal the TLR-4 interactome upon treatment of lipopolysaccharides and statin drug in HA-TLR-4 transfected HEK293 cells. A total of 712 differentially expressed proteins were identified and quantified in this study. Selected candidate proteins, macrophage myristoylated alanine-rich C kinase substrate, and creatine kinase, exclusively identified in the treatment of statin along with cross-linkers, were further validated by immunoblotting.