A unique surface membrane anchored purine-salvage enzyme is conserved among a group of primitive eukaryotic human pathogens.
Previously, we isolated and characterized the gene encoding the 3'-Nucleotidase/Nuclease (Ld3'NT/NU) from the human pathogen, Leishmania donovani. This unique cell surface enzyme has been shown to be involved in the salvage of host-derived purines, which are essential for the survival of this important protozoan parasite. In this report, we assessed whether the 3'-Nucleotidase/Nuclease was conserved amongst other pathogenic Leishmania and related trypanosomatid parasites. Results of pulsed field gel electrophoresis and Southern blotting showed that a Ld3'NT/NU gene homolog was present in each of the visceral and cutaneous Leishmania species tested (i.e. isolates of L. donovani, L. infantum, L. tropica, L. major and L. mexicana, respectively). Further, results of colorimetric assays using 3'-adenosine monophosphate as substrate demonstrated that each of these organisms also expressed significant levels of 3'-nucleotidase enzyme activity. In addition, we showed that a Ld3'NT/NU gene homolog was expressed in each of these Leishmania species as a > 40 kDa 3'-nucleotidase enzyme activity. A Ld3'NT/NU gene homolog was also identified in two Crithidia species (C. fasciculata and C. luciliae) and Leptomonas seymouri but was only marginally detectable in Trypanosoma brucei, Trypanosoma cruzi and Phytomonas serpens. Cumulatively, results of this study showed that an Ld3'NT/NU homolog was conserved amongst pathogenic Leishmania sp. which suggests that this enzyme must play an critical role in purine salvage for all members of this group of human pathogens.
Debrabant
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A
Division of Emerging and Transfusion Transmitted Diseases, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD, USA.
Bastien
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Dwyer
D M
DM
D000818:Animals; D015139:Blotting, Southern; D015153:Blotting, Western; D016054:DNA, Protozoan / Q000032:analysis; D004587:Electrophoresis, Agar Gel; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes; D006801:Humans; D007893:Leishmania donovani / Q000737:chemistry*; D009708:Nucleotidases / Q000737:chemistry / Q000302:isolation & purification; D009841:Oligonucleotides / Q000378:metabolism; D011485:Protein Binding; D011687:Purines / Q000737:chemistry; D016053:RNA, Protozoan / Q000032:analysis; D014345:Trypanosoma / Q000737:chemistry*
D016054:DNA, Protozoan; D000939:Epitopes; D009841:Oligonucleotides; D011687:Purines; D016053:RNA, Protozoan; D009708:Nucleotidases; C040470:3'-nucleotidase