Leishmania (Viannia) braziliensis nucleoside triphosphate diphosphohydrolase (NTPDase 1): Localization and in vitro inhibition of promastigotes growth by polyclonal antibodies |
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| Authors: | Gabriane Nascimento Porcino Cristiane Carvalho-Campos Ana Carolina Ribeiro Gomes Maia Michelle Lima Detoni Priscila Faria-Pinto Elaine Soares Coimbra Marcos José Marques Maria Aparecida Juliano Luiz Juliano Vanessa Álvaro Diniz Suzana Corte-Real Eveline Gomes Vasconcelos |
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| Institution: | 1. 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;2. Departamento de Ciências Biológicas, Instituto de Ciências Biomédicas, Universidade Federal de Alfenas, Alfenas, MG, Brazil;3. Departamento de Biofísica, Escola Paulista de Medicina, Universidade Federal de São Paulo, São Paulo, SP, Brazil;4. Laboratório de Biologia Estrutural, Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, RJ, Brazil |
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| 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. |
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