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Identification of the first pyrimidine nucleobase transporter in Leishmania: similarities with the Trypanosoma brucei U1 transporter and antileishmanial activity of uracil analogues

Published online by Cambridge University Press:  27 October 2004

I. G. PAPAGEORGIOU ,
L. YAKOB ,
M. I. AL SALABI ,
G. DIALLINAS ,
K. P. SOTERIADOU  and
H. P. DE KONING
I. G. PAPAGEORGIOU
Affiliation:
Department of Biochemistry, Hellenic Pasteur Institute, 127 Vassilissis Sophias, 115 21 Athens, Greece Department of Botany, Faculty of Biology, University of Athens, Panepistimioupolis 15781 Athens, Greece
L. YAKOB
Affiliation:
Institute of Biomedical and Life Sciences, Division of Infection and Immunity, University of Glasgow, Glasgow G12 8QQ, UK
M. I. AL SALABI
Affiliation:
Institute of Biomedical and Life Sciences, Division of Infection and Immunity, University of Glasgow, Glasgow G12 8QQ, UK
G. DIALLINAS
Affiliation:
Department of Botany, Faculty of Biology, University of Athens, Panepistimioupolis 15781 Athens, Greece
K. P. SOTERIADOU
Affiliation:
Department of Biochemistry, Hellenic Pasteur Institute, 127 Vassilissis Sophias, 115 21 Athens, Greece
H. P. DE KONING
Affiliation:
Institute of Biomedical and Life Sciences, Division of Infection and Immunity, University of Glasgow, Glasgow G12 8QQ, UK
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Abstract

While purine transport has been widely studied in protozoa, almost nothing is known about their capacity to salvage pyrimidines. Here, we report a Leishmania major transporter with high affinity for uracil (Km=0·32±0·07 μM) which we designated LmU1. This transporter displayed a high degree of specificity, as it had virtually no affinity for cytosine, thymine or purine nucleobases, nor did it transport pyrimidine nucleosides. Highest affinity was for 5-fluorouracil. The results show that the permeant binding site of LmU1 interacts strongly with the keto groups of uracil, as shown by a low affinity for 2-thio- and 4-thiouracil. LmU1 appears to further bind uracil through a weak hydrogen bond with N(1)H of the pyrimidine ring in addition to a stronger H-bond with N(3)H. Substrate binding and selectivity were strikingly similar to that of the U1 transporter in the related kinetoplastid Trypanosoma brucei. Uracil analogues likely to be transported by LmU1 were also screened for antileishmanial activity, with 5-fluorouracil displaying strong activity against promastigotes and intracellular amastigotes. Overall, the results show that, like purine nucleobase transport, pyrimidine nucleobase transport function is very similar in L. major and T. brucei insect forms.

Keywords

Leishmania majorTrypanosoma bruceipyrimidine transporterspurine transportersuracil transporter5-fluorouracil
Type
Research Article
Information
Parasitology , Volume 130 , Issue 3 , March 2005 , pp. 275 - 283
Copyright
2005 Cambridge University Press

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