4DL8: Crystal structure of Trypanosoma brucei dUTPase with dUMP, planar [AlF3-OPO3] transition state analogue, Mg2+, and Na+

Citation:
Abstract
The Tritryps Trypanosoma brucei, Trypanosoma cruzi and Leishmania donovani are responsible for great morbidity and mortality in developing countries. Their dimeric dUTPases are members of the all-alpha NTP pyrophosphohydrolase family and represent promising drug targets due to their essential nature and markedly different structural and biochemical properties compared with the trimeric human enzyme. In the present paper we describe the structure of the T. brucei enzyme in open and closed conformations. Furthermore, we probe the reaction mechanism through the binding of transition state mimics both in solution and in the crystal. 31P-NMR and tryptophan fluorescence quenching in the presence of AlF3 and MgF3- identified which phosphate is subject to nucleophilic attack by a water molecule. The structures in complex with two transition state analogues confirm that the nucleophilic attack occurs on the beta-phosphate in contrast with the alpha-phosphate in the trimeric enzymes. These results establish the structural basis of catalysis of these important housekeeping enzymes and has ramifications for the wider all-alpha NTP pyrophosphohydrolase family.
PDB ID: 4DL8Download
MMDB ID: 112394
PDB Deposition Date: 2012/2/6
Updated in MMDB: 2017/11
Experimental Method:
x-ray diffraction
Resolution: 1.698  Å
Source Organism:
Similar Structures:
Biological Unit for 4DL8: dimeric; determined by author and by software (PISA)
Molecular Components in 4DL8
Label Count Molecule
Proteins (2 molecules)
2
Deoxyuridine Triphosphatase
Molecule annotation
Chemicals (12 molecules)
1
2
2
2
3
2
4
4
5
2
* Click molecule labels to explore molecular sequence information.

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