Host-pathogen interaction profiling using self-assembling human protein arrays

Journal of Proteome Research
Xiaobo YuMatthias P Machner

Abstract

Host-pathogen protein interactions are fundamental to every microbial infection, yet their identification has remained challenging due to the lack of simple detection tools that avoid abundance biases while providing an open format for experimental modifications. Here, we applied the Nucleic Acid-Programmable Protein Array and a HaloTag-Halo ligand detection system to determine the interaction network of Legionella pneumophila effectors (SidM and LidA) with 10 000 unique human proteins. We identified known targets of these L. pneumophila proteins and potentially novel interaction candidates. In addition, we applied our Click chemistry-based NAPPA platform to identify the substrates for SidM, an effector with an adenylyl transferase domain that catalyzes AMPylation (adenylylation), the covalent addition of adenosine monophosphate (AMP). We confirmed a subset of the novel SidM and LidA targets in independent in vitro pull-down and in vivo cell-based assays, and provided further insight into how these effectors may discriminate between different host Rab GTPases. Our method circumvents the purification of thousands of human and pathogen proteins, and does not require antibodies against or prelabeling of query proteins. This system...Continue Reading

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Citations

Feb 7, 2017·Molecular & Cellular Proteomics : MCP·Todd M Greco, Ileana M Cristea
Mar 30, 2018·FEMS Microbiology Reviews·Eric MartinezMatteo Bonazzi
Dec 1, 2017·Frontiers in Cellular and Infection Microbiology·Soma Ghosh, Tamara J O'Connor
Dec 8, 2017·Expert Review of Proteomics·Xiaobo YuJoshua LaBaer
Aug 9, 2017·Expert Review of Proteomics·Alla GagarinovaMohan Babu
Feb 13, 2018·BMC Microbiology·Xiaobo YuM Ramona Neunuebel
Apr 19, 2020·Molecular & Cellular Proteomics : MCP·Guan-Da SyuHeng Zhu
Aug 31, 2019·Expert Review of Proteomics·Huan QiSheng-Ce Tao
Mar 23, 2017·Journal of Immunology Research·Nadia Martinez-Martin
May 11, 2018·Proteomics·Megan H Wright
Apr 30, 2019·Frontiers in Physiology·Joseph Gillen, Aleksandra Nita-Lazar
Jan 13, 2016·The Journal of Biological Chemistry·Ernest C SoGad Frankel
Jan 1, 2015·Microarrays·Paula DíezManuel Fuentes
Jan 17, 2021·Molecular & Cellular Proteomics : MCP·Guan-Da SyuHeng Zhu
Sep 4, 2017·International Journal of Medical Microbiology : IJMM·Bernhard SteinerHubert Hilbi
Mar 4, 2017·Current Protocols in Cell Biology·Yanyang TangJoshua LaBaer

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