PDBsum entry 2wzf

Transferase

PDB id: 2wzf

Contents

Protein chain

515 a.a.

Ligands

UDP

BGC

Metals

_MN

Waters ×316

PDB id:

2wzf

Name:

Transferase

Title:

Legionella pneumophila glucosyltransferase crystal structure

Structure:

Glucosyltransferase. Chain: a. Engineered: yes

Source:

Legionella pneumophila. Organism_taxid: 446. Expressed in: escherichia coli. Expression_system_taxid: 469008.

Resolution:

2.10Å R-factor: 0.189 R-free: 0.254

Authors:

R.Hurtado-Guerrero,T.Zusman,S.Pathak,A.F.M.Ibrahim,S.Shepherd, A.Prescott,G.Segal,D.M.F.Van Aalten

Key ref:

R.Hurtado-Guerrero et al. (2010). Molecular mechanism of elongation factor 1A inhibition by a Legionella pneumophila glycosyltransferase. Biochem J, 426, 281-292. PubMed id: 20030628

Date:

29-Nov-09 Release date: 08-Dec-09

Protein chain

Q5ZVS2  (Q5ZVS2_LEGPH) -  Glucosyltransferase Lgt1 from Legionella pneumophila subsp. pneumophila (strain Philadelphia 1 / ATCC 33152 / DSM 7513)

Seq: 525 a.a.

Struc: 515 a.a.

Enzyme reactions

• Enzyme class: E.C.?

Biochem J 426:281-292 (2010)

PubMed id: 20030628

Molecular mechanism of elongation factor 1A inhibition by a Legionella pneumophila glycosyltransferase.

R.Hurtado-Guerrero, T.Zusman, S.Pathak, A.F.Ibrahim, S.Shepherd, A.Prescott, G.Segal, D.M.van Aalten.

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.