CobB interacting proteins identified by SPIDER assay

To assess whether SPIDER could detect transient interactions such as enzymes and their substrates, we examined the interactome of the E. coli protein deacetylase CobB. As the only member of the Sir2 family of deacetylases in E. coli, CobB is known to play a role in many different pathways but their interactors is still incompletely known. Here we applied SPIDER assay and a SILAC-based mass spectrometry strategy to capture and identify CobB Substrates. Biotinylated CobB was incubated with E. coli total lysate labeled with heavy stable isotope. As a control, free biotin was incubated with E. coli total lysate labeled with light stable isotope.