TY - JOUR AB - The Dot/Icm type IV secretion system (T4SS) is essential for the pathogenesis of Legionella species and translocates a multitude of effector proteins into host cells. The identification of host cell targets of these effectors is often critical to unravel their roles in controlling the host. Here we describe a method to characterize the protein complexes associated with effectors in infected host cells. To achieve this, Legionella expressing an effector of interest fused to a Bio-tag, a combination of hexahistidine tags and a specific recognition sequence for the biotin ligase BirA, are used to infect host cells expressing BirA, which leads to biotinylation of the translocated effector. Following chemical cross-linking, effector interactomes are isolated by tandem affinity purification employing metal affinity and NeutrAvidin resins and identified by western blotting or mass spectrometry. AU - So,EC AU - Mousnier,A AU - Frankel,G AU - Schroeder,GN DO - 10.1007/978-1-4939-9048-1_19 EP - 303 PY - 2019/// SP - 289 TI - Determination of In Vivo Interactomes of Dot/Icm Type IV Secretion System Effectors by Tandem Affinity Purification. T2 - Methods Mol Biol UR - http://dx.doi.org/10.1007/978-1-4939-9048-1_19 UR - https://www.ncbi.nlm.nih.gov/pubmed/30694500 VL - 1921 ER -