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Legionella pp 277-287 | Cite as

Purification and Analysis of Effector Glucosyltransferase Lgt1 from Legionella pneumophila

  • Nadezhda Levanova
  • Irina Tabakova
  • Thomas Jank
  • Yury BelyiEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1921)

Abstract

Legionella pneumophila is a facultative intracellular pathogen responsible for legionellosis, a severe lung disease in humans. This bacterium uses a type 4b secretion system to deliver various effector proteins into the cytoplasm of a eukaryotic target cell. Among those is the glucosyltransferase Lgt1. This effector modifies serine-53 in eukaryotic elongation factor 1A (eEF1A) by mono-O-glucosylation. Modification of eEF1A results in inhibition of protein synthesis and death of the eukaryotic cell, processes which are thought to contribute to Legionella infection. Here we describe a protocol for isolation of the glucosyltransferase Lgt1 from L. pneumophila culture followed by assaying its enzymatic activity using 14C-UDP-glucose autoradiography.

Key words

Legionella Glucosyltransferase Lgt1 Purification UDP-[14C]glucose Autoradiography eEF1A In vitro glucosylation Protein synthesis 

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  • Nadezhda Levanova
    • 1
  • Irina Tabakova
    • 2
  • Thomas Jank
    • 1
  • Yury Belyi
    • 2
    Email author
  1. 1.Faculty of MedicineInstitute for Experimental and Clinical Pharmacology and Toxicology, University of FreiburgFreiburgGermany
  2. 2.Gamaleya Research CentreMoscowRussia

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