In this section
@article{So:2019:10.1007/978-1-4939-9048-1_19,
author = {So, EC and Mousnier, A and Frankel, G and Schroeder, GN},
doi = {10.1007/978-1-4939-9048-1_19},
journal = {Methods Mol Biol},
pages = {289--303},
title = {Determination of In Vivo Interactomes of Dot/Icm Type IV Secretion System Effectors by Tandem Affinity Purification.},
url = {http://dx.doi.org/10.1007/978-1-4939-9048-1_19},
volume = {1921},
year = {2019}
}
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 -
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