Leishmania (Viannia) braziliensis nucleoside triphosphate diphosphohydrolase (NTPDase 1): Localization and in vitro inhibition of promastigotes growth by polyclonal antibodies

Leishmania (Viannia) braziliensis nucleoside triphosphate diphosphohydrolase (NTPDase 1): Localization and in vitro inhibition of promastigotes growth by polyclonal antibodies

Author Porcino, Gabriane Nascimento Google Scholar
Carvalho-Campos, Cristiane Google Scholar
Ribeiro Gomes Maia, Ana Carolina Google Scholar
Detoni, Michelle Lima Google Scholar
Faria-Pinto, Priscila Google Scholar
Coimbra, Elaine Soares Google Scholar
Marques, Marcos Jose Google Scholar
Juliano, Maria Aparecida Autor UNIFESP Google Scholar
Juliano, Luiz Autor UNIFESP Google Scholar
Diniz, Vanessa Alvaro Google Scholar
Corte-Real, Suzana Google Scholar
Vasconcelos, Eveline Gomes Google Scholar
Institution Univ Fed Juiz de Fora
Univ Fed Alfenas
Fiocruz MS
Universidade Federal de São Paulo (UNIFESP)
Abstract Nucleoside triphosphate diphosphohydrolase (NTPDase) activity was recently characterized in Leishmania (Viannia) braziliensis promastigotes (Lb), and an antigenic conserved domain (r82-121) from the specific NTPDase 1 isoform was identified. in this work, mouse polyclonal antibodies produced against two synthetic peptides derived from this domain (LbB1LJ, r82-103; LbB2LJ, r102-121) were used. the anti-LbB1LJ or anti-LbB2LJ antibodies were immobilized on protein A-sepharose and immunoprecipitated the NTPDase 1 of 48 kDa and depleted approximately 40% of the phosphohydrolytic activity from detergent-homogenized Lb preparation. Ultrastructural immunocytochemical microscopy identified the NTPDase 1 on the parasite surface and in its subcellular cytoplasmic vesicles, mitochondria, kinetoplast and nucleus. the ATPase and ADPase activities of detergent-homogenized Lb preparation were partially inhibited by anti-LbB1LJ antibody (43-79%), which was more effective than that inhibition (18-47%) by anti-LbB2LJ antibody. in addition, the immune serum anti-LbB1LJ (67%) or anti-LbB2LJ (33%) was cytotoxic, significantly reducing the promastigotes growth in vitro. the results appoint the conserved domain from the L braziliensis NTPDase as an important target for inhibitor design and the potential application of these biomolecules in experimental protocols of disease control. (c) 2012 Elsevier Inc. All rights reserved.
Keywords Apyrase
ATP diphosphohydrolase
NTPase
Peptide
Leishmaniasis
Immunocytochemical
Language English
Sponsor Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG)
PROQUALI/UFJF
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Grant number FAPEMIG: CBB-APQ-01384-09
FAPEMIG: CBB-APQ 00754-09
Date 2012-10-01
Published in Experimental Parasitology. San Diego: Academic Press Inc Elsevier Science, v. 132, n. 2, p. 293-299, 2012.
ISSN 0014-4894 (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 293-299
Origin http://dx.doi.org/10.1016/j.exppara.2012.08.009
Access rights Open access Open Access
Type Article
Web of Science ID WOS:000309737900027
URI http://repositorio.unifesp.br/handle/11600/35295

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