title: Leishmania (Viannia) braziliensis nucleoside triphosphate diphosphohydrolase (NTPDase 1): localization and in vitro inhibition of promastigotes growth by polyclonal antibodies.
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.
authors.affiliation: Departamento de Bioquímica, Pós-Graduação em Imunologia e DIP/Genética e Biotecnologia, Instituto de Ciências Biológicas, Universidade Federal de Juiz de Fora, Juiz de Fora, MG, Brazil.
authors.forename: Gabriane Nascimento
authors.initials: GN
authors.lastname: Porcino
authors.forename: Cristiane
authors.initials: C
authors.lastname: Carvalho-Campos
authors.forename: Ana Carolina Ribeiro Gomes
authors.initials: AC
authors.lastname: Maia
authors.forename: Michelle Lima
authors.initials: ML
authors.lastname: Detoni
authors.forename: Priscila
authors.initials: P
authors.lastname: Faria-Pinto
authors.forename: Elaine Soares
authors.initials: ES
authors.lastname: Coimbra
authors.forename: Marcos José
authors.initials: MJ
authors.lastname: Marques
authors.forename: Maria Aparecida
authors.initials: MA
authors.lastname: Juliano
authors.forename: Luiz
authors.initials: L
authors.lastname: Juliano
authors.forename: Vanessa Álvaro
authors.initials: VÁ
authors.lastname: Diniz
authors.forename: Suzana
authors.initials: S
authors.lastname: Corte-Real
authors.forename: Eveline Gomes
authors.initials: EG
authors.lastname: Vasconcelos
mesh_terms: D000818:Animals; D055910:Antibodies, Immobilized / Q000276:immunology; D000913:Antibodies, Protozoan / Q000276:immunology*; D015703:Antigens, CD / Q000032:analysis* / Q000276:immunology; D001081:Apyrase / Q000032:analysis* / Q000037:antagonists & inhibitors / Q000276:immunology; D015153:Blotting, Western; D047468:Immunoprecipitation; D007527:Isoenzymes / Q000032:analysis / Q000276:immunology; D007892:Leishmania braziliensis / Q000201:enzymology* / Q000254:growth & development / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D016253:Microscopy, Immunoelectron; D011817:Rabbits
chemical_list: D055910:Antibodies, Immobilized; D000913:Antibodies, Protozoan; D015703:Antigens, CD; D007527:Isoenzymes; D001081:Apyrase; C067869:CD39 antigen