Leishmania amazonensis: Biological and biochemical characterization of ecto-nucleoside triphosphate diphosphohydrolase activities.
The presence of Leishmania amazonensis ecto-nucleoside triphosphate triphosphohydrolase activities was demonstrated using antibodies against different NTPDase members by Western blotting, flow cytometry, and immunoelectron microscopy analysis. Living promastigote cells sequentially hydrolyzed the ATP molecule generating ADP, AMP, and adenosine, indicating that this surface enzyme may play a role in the salvage of purines from the extracellular medium. The L. amazonensis ecto-NTPDase activities were insensitive to Triton X-100, but they were enhanced by divalent cations, such as Mg(2+). In addition, the ecto-NTPDase activities decreased with time for 96 h when promastigotes were grown in vitro. On the other hand, these activities increased considerably when measured in living amastigote forms. Furthermore, the treatment with adenosine, a mediator of several relevant biological phenomena, induced a decrease in the reactivity with anti-CD39 antibody, raised against mammalian E-NTPDase, probably because of down regulation in the L. amazonensis ecto-NTPDase expression. Also, adenosine and anti-NTPDase antibodies induced a significant diminishing in the interaction between promastigotes of L. amazonensis and mouse peritoneal macrophages.
Pinheiro
Carla M
CM
Instituto de Bioquímica Médica (IBqM), Universidade Federal do Rio de Janeiro (UFRJ), Centro de Ciências da Saúde (CCS), Bloco H, Cidade Universitária, Ilha do Fundão, 21541-590 Rio de Janeiro, Brazil.
Martins-Duarte
Erica S
ES
Ferraro
Rodrigo B
RB
Fonseca de Souza
André Luíz
AL
Gomes
Marta T
MT
Lopes
Angela H C S
AH
Vannier-Santos
Marcos A
MA
Santos
André L S
AL
Meyer-Fernandes
José R
JR
D000244:Adenosine Diphosphate / Q000378:metabolism; D000249:Adenosine Monophosphate / Q000378:metabolism; D000251:Adenosine Triphosphatases / Q000378:metabolism*; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D001081:Apyrase / Q000378:metabolism; D015153:Blotting, Western; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007894:Leishmania mexicana / Q000201:enzymology*; D016773:Leishmaniasis, Cutaneous / Q000469:parasitology; D017737:Macrophages, Peritoneal / Q000469:parasitology; D051379:Mice; D016253:Microscopy, Immunoelectron; D011755:Pyrophosphatases / Q000378:metabolism
D015703:Antigens, CD; D000249:Adenosine Monophosphate; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D000251:Adenosine Triphosphatases; D011755:Pyrophosphatases; C027201:ectoATPase; C439652:nucleoside-triphosphate diphosphohydrolase 3; D001081:Apyrase; C067869:CD39 antigen