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

Xiaobo Yu, Kimberly B. Decker, Kristi Barker, M. Ramona Neunuebel, Justin Saul, Morgan Graves, Nathan Westcott, Howard Hang, Joshua LaBaer, Ji Qiu, Matthias P. Machner

Research output: Contribution to journalArticle

27 Scopus citations

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 is amenable to high-throughput analysis of effectors from a wide variety of human pathogens that may bind to and/or post-translationally modify targets within the human proteome.

Original languageEnglish (US)
Pages (from-to)1920-1936
Number of pages17
JournalJournal of Proteome Research
Volume14
Issue number4
DOIs
StatePublished - Apr 3 2015

Keywords

  • AMPylation
  • LidA
  • Rab1
  • SidM
  • interactome
  • nucleic acid programmable protein array (NAPPA)

ASJC Scopus subject areas

  • Biochemistry
  • Chemistry(all)

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  • Cite this

    Yu, X., Decker, K. B., Barker, K., Neunuebel, M. R., Saul, J., Graves, M., Westcott, N., Hang, H., LaBaer, J., Qiu, J., & Machner, M. P. (2015). Host-pathogen interaction profiling using self-assembling human protein arrays. Journal of Proteome Research, 14(4), 1920-1936. https://doi.org/10.1021/pr5013015