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

Detalhes bibliográficos
Autor(a) principal: Porcino, Gabriane Nascimento
Data de Publicação: 2012
Outros Autores: Carvalho-Campos, Cristiane, Ribeiro Gomes Maia, Ana Carolina, Detoni, Michelle Lima, Faria-Pinto, Priscila, Coimbra, Elaine Soares, Marques, Marcos Jose, Juliano, Maria Aparecida [UNIFESP], Juliano, Luiz [UNIFESP], Diniz, Vanessa Alvaro, Corte-Real, Suzana, Vasconcelos, Eveline Gomes
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNIFESP
Texto Completo: http://dx.doi.org/10.1016/j.exppara.2012.08.009
http://repositorio.unifesp.br/handle/11600/35295
Resumo: 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.
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spelling Leishmania (Viannia) braziliensis nucleoside triphosphate diphosphohydrolase (NTPDase 1): Localization and in vitro inhibition of promastigotes growth by polyclonal antibodiesApyraseATP diphosphohydrolaseNTPasePeptideLeishmaniasisImmunocytochemicalNucleoside 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.Univ Fed Juiz de Fora, Inst Ciencias Biol, Dept Bioquim, BR-36036900 Juiz de Fora, MG, BrazilUniv Fed Alfenas, Inst Ciencias Biomed, Dept Ciencias Biol, Alfenas, MG, BrazilFiocruz MS, Inst Oswaldo Cruz, Lab Biol Estrut, Rio de Janeiro, RJ, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Biofis, São Paulo, BrazilUniversidade Federal de São Paulo, Escola Paulista Med, Dept Biofis, São Paulo, BrazilWeb of ScienceFundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG)PROQUALI/UFJFCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)FAPEMIG: CBB-APQ-01384-09FAPEMIG: CBB-APQ 00754-09Elsevier B.V.Univ Fed Juiz de ForaUniv Fed AlfenasFiocruz MSUniversidade Federal de São Paulo (UNIFESP)Porcino, Gabriane NascimentoCarvalho-Campos, CristianeRibeiro Gomes Maia, Ana CarolinaDetoni, Michelle LimaFaria-Pinto, PriscilaCoimbra, Elaine SoaresMarques, Marcos JoseJuliano, Maria Aparecida [UNIFESP]Juliano, Luiz [UNIFESP]Diniz, Vanessa AlvaroCorte-Real, SuzanaVasconcelos, Eveline Gomes2016-01-24T14:27:43Z2016-01-24T14:27:43Z2012-10-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersion293-299application/pdfhttp://dx.doi.org/10.1016/j.exppara.2012.08.009Experimental Parasitology. San Diego: Academic Press Inc Elsevier Science, v. 132, n. 2, p. 293-299, 2012.10.1016/j.exppara.2012.08.009WOS000309737900027.pdf0014-4894http://repositorio.unifesp.br/handle/11600/35295WOS:000309737900027engExperimental Parasitologyinfo:eu-repo/semantics/openAccesshttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policyreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESP2024-08-01T02:48:00Zoai:repositorio.unifesp.br/:11600/35295Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestbiblioteca.csp@unifesp.bropendoar:34652024-08-01T02:48Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false
dc.title.none.fl_str_mv Leishmania (Viannia) braziliensis nucleoside triphosphate diphosphohydrolase (NTPDase 1): Localization and in vitro inhibition of promastigotes growth by polyclonal antibodies
title Leishmania (Viannia) braziliensis nucleoside triphosphate diphosphohydrolase (NTPDase 1): Localization and in vitro inhibition of promastigotes growth by polyclonal antibodies
spellingShingle Leishmania (Viannia) braziliensis nucleoside triphosphate diphosphohydrolase (NTPDase 1): Localization and in vitro inhibition of promastigotes growth by polyclonal antibodies
Porcino, Gabriane Nascimento
Apyrase
ATP diphosphohydrolase
NTPase
Peptide
Leishmaniasis
Immunocytochemical
title_short Leishmania (Viannia) braziliensis nucleoside triphosphate diphosphohydrolase (NTPDase 1): Localization and in vitro inhibition of promastigotes growth by polyclonal antibodies
title_full Leishmania (Viannia) braziliensis nucleoside triphosphate diphosphohydrolase (NTPDase 1): Localization and in vitro inhibition of promastigotes growth by polyclonal antibodies
title_fullStr Leishmania (Viannia) braziliensis nucleoside triphosphate diphosphohydrolase (NTPDase 1): Localization and in vitro inhibition of promastigotes growth by polyclonal antibodies
title_full_unstemmed Leishmania (Viannia) braziliensis nucleoside triphosphate diphosphohydrolase (NTPDase 1): Localization and in vitro inhibition of promastigotes growth by polyclonal antibodies
title_sort Leishmania (Viannia) braziliensis nucleoside triphosphate diphosphohydrolase (NTPDase 1): Localization and in vitro inhibition of promastigotes growth by polyclonal antibodies
author Porcino, Gabriane Nascimento
author_facet Porcino, Gabriane Nascimento
Carvalho-Campos, Cristiane
Ribeiro Gomes Maia, Ana Carolina
Detoni, Michelle Lima
Faria-Pinto, Priscila
Coimbra, Elaine Soares
Marques, Marcos Jose
Juliano, Maria Aparecida [UNIFESP]
Juliano, Luiz [UNIFESP]
Diniz, Vanessa Alvaro
Corte-Real, Suzana
Vasconcelos, Eveline Gomes
author_role author
author2 Carvalho-Campos, Cristiane
Ribeiro Gomes Maia, Ana Carolina
Detoni, Michelle Lima
Faria-Pinto, Priscila
Coimbra, Elaine Soares
Marques, Marcos Jose
Juliano, Maria Aparecida [UNIFESP]
Juliano, Luiz [UNIFESP]
Diniz, Vanessa Alvaro
Corte-Real, Suzana
Vasconcelos, Eveline Gomes
author2_role author
author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Univ Fed Juiz de Fora
Univ Fed Alfenas
Fiocruz MS
Universidade Federal de São Paulo (UNIFESP)
dc.contributor.author.fl_str_mv Porcino, Gabriane Nascimento
Carvalho-Campos, Cristiane
Ribeiro Gomes Maia, Ana Carolina
Detoni, Michelle Lima
Faria-Pinto, Priscila
Coimbra, Elaine Soares
Marques, Marcos Jose
Juliano, Maria Aparecida [UNIFESP]
Juliano, Luiz [UNIFESP]
Diniz, Vanessa Alvaro
Corte-Real, Suzana
Vasconcelos, Eveline Gomes
dc.subject.por.fl_str_mv Apyrase
ATP diphosphohydrolase
NTPase
Peptide
Leishmaniasis
Immunocytochemical
topic Apyrase
ATP diphosphohydrolase
NTPase
Peptide
Leishmaniasis
Immunocytochemical
description 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.
publishDate 2012
dc.date.none.fl_str_mv 2012-10-01
2016-01-24T14:27:43Z
2016-01-24T14:27:43Z
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://dx.doi.org/10.1016/j.exppara.2012.08.009
Experimental Parasitology. San Diego: Academic Press Inc Elsevier Science, v. 132, n. 2, p. 293-299, 2012.
10.1016/j.exppara.2012.08.009
WOS000309737900027.pdf
0014-4894
http://repositorio.unifesp.br/handle/11600/35295
WOS:000309737900027
url http://dx.doi.org/10.1016/j.exppara.2012.08.009
http://repositorio.unifesp.br/handle/11600/35295
identifier_str_mv Experimental Parasitology. San Diego: Academic Press Inc Elsevier Science, v. 132, n. 2, p. 293-299, 2012.
10.1016/j.exppara.2012.08.009
WOS000309737900027.pdf
0014-4894
WOS:000309737900027
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Experimental Parasitology
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
http://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
eu_rights_str_mv openAccess
rights_invalid_str_mv http://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dc.format.none.fl_str_mv 293-299
application/pdf
dc.publisher.none.fl_str_mv Elsevier B.V.
publisher.none.fl_str_mv Elsevier B.V.
dc.source.none.fl_str_mv reponame:Repositório Institucional da UNIFESP
instname:Universidade Federal de São Paulo (UNIFESP)
instacron:UNIFESP
instname_str Universidade Federal de São Paulo (UNIFESP)
instacron_str UNIFESP
institution UNIFESP
reponame_str Repositório Institucional da UNIFESP
collection Repositório Institucional da UNIFESP
repository.name.fl_str_mv Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)
repository.mail.fl_str_mv biblioteca.csp@unifesp.br
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