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Published on: November 22, 2013
Purification and Analysis of Effector Glucosyltransferase Lgt1 from Legionella pneumophila
Nadezhda Levanova1, Irina Tabakova2, Thomas Jank1
1Faculty of Medicine, Institute for Experimental and Clinical Pharmacology and Toxicology, University of Freiburg, Freiburg, Germany.
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.
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