2WZF

Legionella pneumophila glucosyltransferase crystal structure

Summary for 2WZF

• Entry DOI: 10.2210/pdb2wzf/pdb
• Related: 2WZG
• Descriptor: GLUCOSYLTRANSFERASE, MANGANESE (II) ION, URIDINE-5'-DIPHOSPHATE, ... (5 entities in total)
• Functional Keywords: transferase, elongation factor 1a, virulence factor, glucosyltransferase
• Biological source: LEGIONELLA PNEUMOPHILA
• Total number of polymer chains: 1
• Total formula weight: 60383.32

Authors

Hurtado-Guerrero, R.,Zusman, T.,Pathak, S.,Ibrahim, A.F.M.,Shepherd, S.,Prescott, A.,Segal, G.,van Aalten, D.M.F. (deposition date: 2009-11-29, release date: 2009-12-08, Last modification date: 2023-12-20)

Primary citation

Hurtado-Guerrero, R.,Zusman, T.,Pathak, S.,Ibrahim, A.F.M.,Shepherd, S.,Prescott, A.,Segal, G.,Van Aalten, D.M.F.
Molecular Mechanism of Elongation Factor 1A Inhibition by a Legionella Pneumophila Glycosyltransferase.
Biochem.J., 426:281-, 2010

PubMed Abstract: Legionnaires' disease is caused by a lethal colonization of alveolar macrophages with the Gram-negative bacterium Legionella pneumophila. LpGT (L. pneumophila glucosyltransferase; also known as Lgt1) has recently been identified as a virulence factor, shutting down protein synthesis in the human cell by specific glucosylation of EF1A (elongation factor 1A), using an unknown mode of substrate recognition and a retaining mechanism for glycosyl transfer. We have determined the crystal structure of LpGT in complex with substrates, revealing a GT-A fold with two unusual protruding domains. Through structure-guided mutagenesis of LpGT, several residues essential for binding of the UDP-glucose-donor and EF1A-acceptor substrates were identified, which also affected L. pneumophila virulence as demonstrated by microinjection studies. Together, these results suggested that a positively charged EF1A loop binds to a negatively charged conserved groove on the LpGT structure, and that two asparagine residues are essential for catalysis. Furthermore, we showed that two further L. pneumophila glycosyltransferases possessed the conserved UDP-glucose-binding sites and EF1A-binding grooves, and are, like LpGT, translocated into the macrophage through the Icm/Dot (intracellular multiplication/defect in organelle trafficking) system.

PubMed: 20030628
DOI: 10.1042/BJ20091351

Experimental method

X-RAY DIFFRACTION (2.1 Å)