Projects per year
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 Clown protein synthesis in the human cell by specific glucosylation of EF1A (elongation factor 1A), using all Unknown mole 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 oil 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.
Original language | English |
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Pages (from-to) | 281-292 |
Number of pages | 12 |
Journal | Biochemical Journal |
Volume | 426 |
DOIs | |
Publication status | Published - 15 Mar 2010 |
Keywords
- elongation factor 1A (EF1A)
- glucosyl transferase
- Legionella pneumophila
- microinjection
- site-directed mutagenesis
- protein structure
- LARGE CLOSTRIDIAL CYTOTOXINS
- NUCLEOTIDE-EXCHANGE FACTOR
- DIFFICILE TOXIN-B
- HISTO-BLOOD GROUP
- STRUCTURAL BASIS
- CONFORMATIONAL-CHANGES
- CRYSTAL-STRUCTURE
- PROTEIN STRUCTURES
- COXIELLA-BURNETII
- HUMAN MACROPHAGES
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- 2 Finished
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Aref#d: 21559. Molecular Mechanisms of Fungal Cell Wall Assembly (Programme Grant)
van Aalten, D. (Investigator)
1/11/09 → 31/10/14
Project: Research
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Aref#d: 21318. Molecular Mechanisms of O-GlcNAc Signalling (Senior Fellowship Renewal)
van Aalten, D. (Investigator)
1/06/09 → 29/02/16
Project: Research