Characterization of deoxyuridine 5'-triphosphate nucleotidohydrolase from Trypanosoma cruzi.
We report the cloning and kinetic characterization of Trypanosoma cruzi deoxyuridine 5'-triphosphate nucleotidohydrolase (dUTPase) whose coding sequence was isolated by genetic complementation in Escherichia coli. The deduced amino acid sequence was similar to Leishmania major dUTPase although it exhibits an amino acid insertion which is sensitive to protease inactivation. The catalytically active species of the enzyme is a dimer and a detailed kinetic characterization showed that it is highly specific for dUTP and dUDP. The general observation that dUTPases from the Trypanosomatidae differ in sequence, conformation and substrate specificity suggests that a different family of dUTPases exists in certain organisms, which may be exploited as drug targets against infectious diseases.
Bernier-Villamor
Victor
V
Instituto de Parasitologia y Biomedicina Lopez Neyra (CSIC), C/Ventanilla 11, 18001 Granada, Spain.
Camacho
Ana
A
Hidalgo-Zarco
Fernando
F
Pérez
Juana
J
Ruiz-Pérez
Luis M
LM
González-Pacanowska
Dolores
D
D000595:Amino Acid Sequence; D000818:Animals; D004926:Escherichia coli / Q000201:enzymology; D007700:Kinetics; D018320:Leishmania major / Q000201:enzymology; D008324:Mammary Tumor Virus, Mouse / Q000201:enzymology; D008969:Molecular Sequence Data; D011755:Pyrophosphatases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013379:Substrate Specificity; D014349:Trypanosoma cruzi / Q000201:enzymology*
D011994:Recombinant Proteins; D011755:Pyrophosphatases; C020020:dUTP pyrophosphatase