This project aims to characterize the functional interaction between the E3 ubiquitin ligase RNF114 and the Toll-pathway signaling kinase TBK1 (TANK-binding kinase 1). TBK1 is a central orchestrator of innate antiviral immunity and inflammatory responses. While RNF114 has been implicated in the regulation of immune signaling, the precise molecular "map" of how it modifies TBK1 remains elusive. This study utilizes Liquid Chromatography-Tandem Mass Spectrometry (LC-MS/MS) to identify specific lysine residues on TBK1 that undergo enhanced ubiquitination in the presence of RNF114.