Measurement of effector protein translocation using phosphorylatable epitope tags and phospho-specific antibodies

Sara Schesser Bartra, Gregory V Plano

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Numerous bacterial pathogens employ specialized protein secretion machineries to directly inject anti-host proteins, termed effector proteins, into eukaryotic cells. Effector proteins carrying small phosphorylatable tags can be used to detect and quantify effector protein injection. Here, we describe the use of the ELKand GSK-tags to detect the translocation of the Y. pestis YopE effector protein into RAW 264.7 macrophagelike cells using immunoblot analysis with phospho-specific antibodies.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages111-119
Number of pages9
Volume1531
DOIs
StatePublished - 2017

Publication series

NameMethods in Molecular Biology
Volume1531
ISSN (Print)10643745

Keywords

  • Effector proteins
  • Phospho-specific antibodies
  • Translocation
  • Type III secretion
  • Type IV secretion
  • Yersinia pestis

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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    Bartra, S. S., & Plano, G. V. (2017). Measurement of effector protein translocation using phosphorylatable epitope tags and phospho-specific antibodies. In Methods in Molecular Biology (Vol. 1531, pp. 111-119). (Methods in Molecular Biology; Vol. 1531). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-6649-3_10