3B82

Structure of the eEF2-ExoA(E546H)-NAD+ complex

Summary for 3B82

• Entry DOI: 10.2210/pdb3b82/pdb
• Related: 1ZM2 1ZM3 1ZM4 1ZM9 3B78 3B8H
• Descriptor: Elongation factor 2, Exotoxin A, NICOTINAMIDE-ADENINE-DINUCLEOTIDE, ... (4 entities in total)
• Functional Keywords: elongation factor, toxin, adp-ribosylation, toxin-substrate complex, cytoplasm, gtp-binding, nucleotide-binding, phosphorylation, protein biosynthesis, rna-binding, rrna-binding, glycosyltransferase, nad, transferase, biosynthetic protein-transferase complex, biosynthetic protein/transferase
• Biological source: Pseudomonas aeruginosa; More
• Cellular location: Cytoplasm: P32324
• Total number of polymer chains: 6
• Total formula weight: 349979.26

Authors

Jorgensen, R.,Merrill, A.R. (deposition date: 2007-10-31, release date: 2008-06-17, Last modification date: 2023-08-30)

Primary citation

Jorgensen, R.,Wang, Y.,Visschedyk, D.,Merrill, A.R.
The nature and character of the transition state for the ADP-ribosyltransferase reaction.
Embo Rep., 9:802-809, 2008

PubMed Abstract: Exotoxin A (ExoA) from Pseudomonas aeruginosa is an important virulence factor that belongs to a class of exotoxins that are secreted by pathogenic bacteria which cause human diseases such as cholera, diphtheria, pneumonia and whooping cough. We present the first crystal structures, to our knowledge, of ExoA in complex with elongation factor 2 (eEF2) and intact NAD(+), which indicate a direct role of two active-site loops in ExoA during the catalytic cycle. One loop moves to form a solvent cover for the active site of the enzyme and reaches towards the target residue (diphthamide) in eEF2 forming an important hydrogen bond. The NAD(+) substrate adopts a conformation remarkably different from that of the NAD(+) analogue, betaTAD, observed in previous structures, and fails to trigger any loop movements. Mutational studies of the two loops in the toxin identify several residues important for catalytic activity, in particular Glu 546 and Arg 551, clearly supporting the new complex structures. On the basis of these data, we propose a transition-state model for the toxin-catalysed reaction.

PubMed: 18583986
DOI: 10.1038/embor.2008.90

Experimental method

X-RAY DIFFRACTION (2.35 Å)