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://repositorio.unifesp.br/handle/11600/35295 http://dx.doi.org/10.1016/j.exppara.2012.08.009 |
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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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 GomesUniv Fed Juiz de ForaUniv Fed AlfenasFiocruz MSUniversidade Federal de São Paulo (UNIFESP)2016-01-24T14:27:43Z2016-01-24T14:27:43Z2012-10-01Experimental Parasitology. San Diego: Academic Press Inc Elsevier Science, v. 132, n. 2, p. 293-299, 2012.0014-4894http://repositorio.unifesp.br/handle/11600/35295http://dx.doi.org/10.1016/j.exppara.2012.08.009WOS000309737900027.pdf10.1016/j.exppara.2012.08.009WOS:000309737900027Nucleoside 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.Fundação de Amparo à Pesquisa do Estado de Minas Gerais (FAPEMIG)PROQUALI/UFJFCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)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, BrazilFAPEMIG: CBB-APQ-01384-09FAPEMIG: CBB-APQ 00754-09Web of Science293-299engElsevier B.V.Experimental Parasitologyhttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policyinfo:eu-repo/semantics/openAccessApyraseATP diphosphohydrolaseNTPasePeptideLeishmaniasisImmunocytochemicalLeishmania (Viannia) braziliensis nucleoside triphosphate diphosphohydrolase (NTPDase 1): Localization and in vitro inhibition of promastigotes growth by polyclonal antibodiesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlereponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESPORIGINALWOS000309737900027.pdfapplication/pdf726309${dspace.ui.url}/bitstream/11600/35295/1/WOS000309737900027.pdfb3868905d222156e9dbc2372db35cff5MD51open accessTEXTWOS000309737900027.pdf.txtWOS000309737900027.pdf.txtExtracted texttext/plain41674${dspace.ui.url}/bitstream/11600/35295/2/WOS000309737900027.pdf.txt35d5097eaddbcfd0010990afd1778303MD52open access11600/352952022-06-02 09:34:41.562open accessoai:repositorio.unifesp.br:11600/35295Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestopendoar:34652022-06-02T12:34:41Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false |
dc.title.en.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.institution.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.eng.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.issued.fl_str_mv |
2012-10-01 |
dc.date.accessioned.fl_str_mv |
2016-01-24T14:27:43Z |
dc.date.available.fl_str_mv |
2016-01-24T14:27:43Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
Experimental Parasitology. San Diego: Academic Press Inc Elsevier Science, v. 132, n. 2, p. 293-299, 2012. |
dc.identifier.uri.fl_str_mv |
http://repositorio.unifesp.br/handle/11600/35295 http://dx.doi.org/10.1016/j.exppara.2012.08.009 |
dc.identifier.issn.none.fl_str_mv |
0014-4894 |
dc.identifier.file.none.fl_str_mv |
WOS000309737900027.pdf |
dc.identifier.doi.none.fl_str_mv |
10.1016/j.exppara.2012.08.009 |
dc.identifier.wos.none.fl_str_mv |
WOS:000309737900027 |
identifier_str_mv |
Experimental Parasitology. San Diego: Academic Press Inc Elsevier Science, v. 132, n. 2, p. 293-299, 2012. 0014-4894 WOS000309737900027.pdf 10.1016/j.exppara.2012.08.009 WOS:000309737900027 |
url |
http://repositorio.unifesp.br/handle/11600/35295 http://dx.doi.org/10.1016/j.exppara.2012.08.009 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.ispartof.none.fl_str_mv |
Experimental Parasitology |
dc.rights.driver.fl_str_mv |
http://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy info:eu-repo/semantics/openAccess |
rights_invalid_str_mv |
http://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
293-299 |
dc.publisher.none.fl_str_mv |
Elsevier B.V. |
publisher.none.fl_str_mv |
Elsevier B.V. |
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Repositório Institucional da UNIFESP |
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Repositório Institucional da UNIFESP |
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