FIGURE 1.
Schematic workflow of isolation and identification of Bio-tagged effector complexes. A549-BirA cells are infected with Legionella strains expressing His6-Bio-tagged effectors. Upon translocation, the Bio-tagged effector (orange oval) is biotinylated by the biotin ligase, BirA. Interactions between the effector and its host targets are stabilized using chemical crosslinking (in red). Subsequent tandem-affinity purification of effector complexes using His6/Ni2+ NTA followed by biotin/Neutravidin enables their composition to be determined using LC-MS/MS.