Leishmania (Viannia) braziliensis nucleoside triphosphate diphosphohydrolase (NTPDase 1): Localization and in vitro inhibition of promastigotes growth by polyclonal antibodies
Autor(a) principal: | |
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Data de Publicação: | 2012 |
Outros Autores: | , , , , , , , , , , |
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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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 |
_version_ |
1814268287640403968 |