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Leishmania infantum amastigote nucleoside triphosphate diphosphohydrolase 1 (NTPDase 1): Its inhibition as a new insight into mode of action of pentamidine.

Experimental parasitology (2019-03-25)
Ana Carolina Ribeiro Gomes Maia, Gabriane Nascimento Porcino, Michelle Lima Detoni, Leonardo Ramos Quellis, Nayara Braga Emídio, Danielle Gomes Marconato, Wagner Faria Messias, Laura Lavorato Soldati, Priscila Faria-Pinto, Priscila Vanessa da Silva Zabala Capriles, Elaine Soares Coimbra, Marcos José Marques, Eveline Gomes Vasconcelos
ABSTRACT

Nucleoside triphosphate diphosphohydrolase (NTPDase) 1 from intracellular amastigotes of Leishmania infantum-infected macrophage was identified by immunocytochemistry and confocal laser scanning microscopy using antibodies that specifically recognize its B-domain. This enzyme was previously characterized in Leishmania promastigote form, and here it is shown to be susceptible to pentamidine isethionate (PEN). In initial assays, this antileishmanial compound (100 μM) reduced 60% phosphohydrolytic activity of promastigotes preparation. An active NTPDase 1 was then isolated by non-denaturing gel electrophoresis, and PEN (10 μM) inhibited 74% and 35% of the ATPase and ADPase activities, respectively, of this pure protein. In addition, PEN 0.1-1 μM inhibited 56% potato apyrase activity, a plant protein that shares high identity with Leishmania NTPDase 1. In contrast, amphotericin B, fluconazole, ketoconazole or allopurinol did not significantly affect phosphohydrolytic activity of either promastigotes preparation or potato apyrase. This work suggests amastigote NTPDase 1 as a new molecular target, and inhibition of its catalytic activity by pentamidine can be part of the mode of action of this drug contributing with the knowledge of its antileishmanial effect.

MATERIALS
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Brand
Product Description

Sigma-Aldrich
L-(−)-Malic acid, ≥95% (titration)
Sigma-Aldrich
Nonaethylene glycol monododecyl ether, nonionic surfactant