Biochemical characterization of a protein tyrosine phosphatase from Trypanosoma cruzi involved in metacyclogenesis and cell invasion
Autor(a) principal: | |
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Data de Publicação: | 2011 |
Outros Autores: | , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNIFESP |
Texto Completo: | http://repositorio.unifesp.br/handle/11600/33701 http://dx.doi.org/10.1016/j.bbrc.2011.04.038 |
Resumo: | Protein tyrosine phosphatases (PTPs) form a large family of enzymes involved in the regulation of numerous cellular functions in eukaryotes. Several protein tyrosine phosphatases have been recently identified in trypanosomatides. Here we report the purification and biochemical characterization of TcPTP1, a protein tyrosine phosphatase from Trypanosoma cruzi, the causing agent of Chagas' disease. the enzyme was cloned and expressed recombinantly in Escherichia coli and purified by Ni-affinity chromatography. Biochemical characterization of recombinant TcPTP1 with the PTP pseudo-substrate pNPP allowed the estimation of a Michaelis-Menten constant K-m of 4.5 mM and a k(cat) of 2.8 s(-1). We were able to demonstrate inhibition of the enzyme by the PTP1b inhibitor BZ3, which on its turn was able to accelerate the differentiation of epimastigotes into metacyclic forms of T. cruzi induced by nutritional stress. Additionally, this compound was able to inhibit by 50% the infectivity of T. cruzi trypomastigotes in a separate cellular assay. in conclusion our results indicate that TcPTP1 is of importance for cellular differentiation and invasivity of this parasite and thus is a valid target for the rational drug design of potential antibiotics directed against T. cruzi. (C) 2011 Elsevier Inc. All rights reserved. |
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Gallo, Gloria [UNIFESP]Ramos, Thiago Cesar Prata [UNIFESP]Tavares, FernandaRocha, Antonio Augusto [UNIFESP]Machi, EmersonSchenkman, Sergio [UNIFESP]Bahia, Diana [UNIFESP]Pesquero, João Bosco [UNIFESP]Wuertele, Martin [UNIFESP]Universidade Federal de São Paulo (UNIFESP)Ctr Biol Mol Estrutural2016-01-24T14:16:45Z2016-01-24T14:16:45Z2011-05-13Biochemical and Biophysical Research Communications. San Diego: Academic Press Inc Elsevier Science, v. 408, n. 3, p. 427-431, 2011.0006-291Xhttp://repositorio.unifesp.br/handle/11600/33701http://dx.doi.org/10.1016/j.bbrc.2011.04.038WOS000290926400012.pdf10.1016/j.bbrc.2011.04.038WOS:000290926400012Protein tyrosine phosphatases (PTPs) form a large family of enzymes involved in the regulation of numerous cellular functions in eukaryotes. Several protein tyrosine phosphatases have been recently identified in trypanosomatides. Here we report the purification and biochemical characterization of TcPTP1, a protein tyrosine phosphatase from Trypanosoma cruzi, the causing agent of Chagas' disease. the enzyme was cloned and expressed recombinantly in Escherichia coli and purified by Ni-affinity chromatography. Biochemical characterization of recombinant TcPTP1 with the PTP pseudo-substrate pNPP allowed the estimation of a Michaelis-Menten constant K-m of 4.5 mM and a k(cat) of 2.8 s(-1). We were able to demonstrate inhibition of the enzyme by the PTP1b inhibitor BZ3, which on its turn was able to accelerate the differentiation of epimastigotes into metacyclic forms of T. cruzi induced by nutritional stress. Additionally, this compound was able to inhibit by 50% the infectivity of T. cruzi trypomastigotes in a separate cellular assay. in conclusion our results indicate that TcPTP1 is of importance for cellular differentiation and invasivity of this parasite and thus is a valid target for the rational drug design of potential antibiotics directed against T. cruzi. (C) 2011 Elsevier Inc. All rights reserved.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Universidade Federal de São Paulo, Dept Ciencia & Tecnol, BR-12231280 Sao Jose Dos Campos, BrazilUniversidade Federal de São Paulo, Dept Biofis, BR-04023062 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Microbiol Imunol & Parasitol, BR-04023062 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Morfol & Genet, BR-04023062 São Paulo, BrazilCtr Biol Mol Estrutural, Lab Nacl Luz Sincrotron, BR-13083100 Campinas, SP, BrazilUniversidade Federal de São Paulo, Dept Ciencia & Tecnol, BR-12231280 Sao Jose Dos Campos, BrazilUniversidade Federal de São Paulo, Dept Biofis, BR-04023062 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Microbiol Imunol & Parasitol, BR-04023062 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Morfol & Genet, BR-04023062 São Paulo, BrazilWeb of Science427-431engElsevier B.V.Biochemical and Biophysical Research Communicationshttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policyinfo:eu-repo/semantics/openAccessProtein tyrosine phosphataseTrypanosoma cruziChagas' diseaseRecombinant expressionBiochemical characterization of a protein tyrosine phosphatase from Trypanosoma cruzi involved in metacyclogenesis and cell invasioninfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlereponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESPORIGINALWOS000290926400012.pdfapplication/pdf435994${dspace.ui.url}/bitstream/11600/33701/1/WOS000290926400012.pdf9dc9b07ff39d604af170cb2858c7917aMD51open accessTEXTWOS000290926400012.pdf.txtWOS000290926400012.pdf.txtExtracted texttext/plain26494${dspace.ui.url}/bitstream/11600/33701/9/WOS000290926400012.pdf.txtdd6177fe9b480a5c3568da4bb0dc152fMD59open accessTHUMBNAILWOS000290926400012.pdf.jpgWOS000290926400012.pdf.jpgIM Thumbnailimage/jpeg7305${dspace.ui.url}/bitstream/11600/33701/11/WOS000290926400012.pdf.jpg289708b0a270aacee2220e9d3b9d9da0MD511open access11600/337012023-06-05 19:06:40.144open accessoai:repositorio.unifesp.br:11600/33701Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestopendoar:34652023-06-05T22:06:40Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false |
dc.title.en.fl_str_mv |
Biochemical characterization of a protein tyrosine phosphatase from Trypanosoma cruzi involved in metacyclogenesis and cell invasion |
title |
Biochemical characterization of a protein tyrosine phosphatase from Trypanosoma cruzi involved in metacyclogenesis and cell invasion |
spellingShingle |
Biochemical characterization of a protein tyrosine phosphatase from Trypanosoma cruzi involved in metacyclogenesis and cell invasion Gallo, Gloria [UNIFESP] Protein tyrosine phosphatase Trypanosoma cruzi Chagas' disease Recombinant expression |
title_short |
Biochemical characterization of a protein tyrosine phosphatase from Trypanosoma cruzi involved in metacyclogenesis and cell invasion |
title_full |
Biochemical characterization of a protein tyrosine phosphatase from Trypanosoma cruzi involved in metacyclogenesis and cell invasion |
title_fullStr |
Biochemical characterization of a protein tyrosine phosphatase from Trypanosoma cruzi involved in metacyclogenesis and cell invasion |
title_full_unstemmed |
Biochemical characterization of a protein tyrosine phosphatase from Trypanosoma cruzi involved in metacyclogenesis and cell invasion |
title_sort |
Biochemical characterization of a protein tyrosine phosphatase from Trypanosoma cruzi involved in metacyclogenesis and cell invasion |
author |
Gallo, Gloria [UNIFESP] |
author_facet |
Gallo, Gloria [UNIFESP] Ramos, Thiago Cesar Prata [UNIFESP] Tavares, Fernanda Rocha, Antonio Augusto [UNIFESP] Machi, Emerson Schenkman, Sergio [UNIFESP] Bahia, Diana [UNIFESP] Pesquero, João Bosco [UNIFESP] Wuertele, Martin [UNIFESP] |
author_role |
author |
author2 |
Ramos, Thiago Cesar Prata [UNIFESP] Tavares, Fernanda Rocha, Antonio Augusto [UNIFESP] Machi, Emerson Schenkman, Sergio [UNIFESP] Bahia, Diana [UNIFESP] Pesquero, João Bosco [UNIFESP] Wuertele, Martin [UNIFESP] |
author2_role |
author author author author author author author author |
dc.contributor.institution.none.fl_str_mv |
Universidade Federal de São Paulo (UNIFESP) Ctr Biol Mol Estrutural |
dc.contributor.author.fl_str_mv |
Gallo, Gloria [UNIFESP] Ramos, Thiago Cesar Prata [UNIFESP] Tavares, Fernanda Rocha, Antonio Augusto [UNIFESP] Machi, Emerson Schenkman, Sergio [UNIFESP] Bahia, Diana [UNIFESP] Pesquero, João Bosco [UNIFESP] Wuertele, Martin [UNIFESP] |
dc.subject.eng.fl_str_mv |
Protein tyrosine phosphatase Trypanosoma cruzi Chagas' disease Recombinant expression |
topic |
Protein tyrosine phosphatase Trypanosoma cruzi Chagas' disease Recombinant expression |
description |
Protein tyrosine phosphatases (PTPs) form a large family of enzymes involved in the regulation of numerous cellular functions in eukaryotes. Several protein tyrosine phosphatases have been recently identified in trypanosomatides. Here we report the purification and biochemical characterization of TcPTP1, a protein tyrosine phosphatase from Trypanosoma cruzi, the causing agent of Chagas' disease. the enzyme was cloned and expressed recombinantly in Escherichia coli and purified by Ni-affinity chromatography. Biochemical characterization of recombinant TcPTP1 with the PTP pseudo-substrate pNPP allowed the estimation of a Michaelis-Menten constant K-m of 4.5 mM and a k(cat) of 2.8 s(-1). We were able to demonstrate inhibition of the enzyme by the PTP1b inhibitor BZ3, which on its turn was able to accelerate the differentiation of epimastigotes into metacyclic forms of T. cruzi induced by nutritional stress. Additionally, this compound was able to inhibit by 50% the infectivity of T. cruzi trypomastigotes in a separate cellular assay. in conclusion our results indicate that TcPTP1 is of importance for cellular differentiation and invasivity of this parasite and thus is a valid target for the rational drug design of potential antibiotics directed against T. cruzi. (C) 2011 Elsevier Inc. All rights reserved. |
publishDate |
2011 |
dc.date.issued.fl_str_mv |
2011-05-13 |
dc.date.accessioned.fl_str_mv |
2016-01-24T14:16:45Z |
dc.date.available.fl_str_mv |
2016-01-24T14:16:45Z |
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 |
Biochemical and Biophysical Research Communications. San Diego: Academic Press Inc Elsevier Science, v. 408, n. 3, p. 427-431, 2011. |
dc.identifier.uri.fl_str_mv |
http://repositorio.unifesp.br/handle/11600/33701 http://dx.doi.org/10.1016/j.bbrc.2011.04.038 |
dc.identifier.issn.none.fl_str_mv |
0006-291X |
dc.identifier.file.none.fl_str_mv |
WOS000290926400012.pdf |
dc.identifier.doi.none.fl_str_mv |
10.1016/j.bbrc.2011.04.038 |
dc.identifier.wos.none.fl_str_mv |
WOS:000290926400012 |
identifier_str_mv |
Biochemical and Biophysical Research Communications. San Diego: Academic Press Inc Elsevier Science, v. 408, n. 3, p. 427-431, 2011. 0006-291X WOS000290926400012.pdf 10.1016/j.bbrc.2011.04.038 WOS:000290926400012 |
url |
http://repositorio.unifesp.br/handle/11600/33701 http://dx.doi.org/10.1016/j.bbrc.2011.04.038 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.ispartof.none.fl_str_mv |
Biochemical and Biophysical Research Communications |
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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 |
427-431 |
dc.publisher.none.fl_str_mv |
Elsevier B.V. |
publisher.none.fl_str_mv |
Elsevier B.V. |
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reponame:Repositório Institucional da UNIFESP instname:Universidade Federal de São Paulo (UNIFESP) instacron:UNIFESP |
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Universidade Federal de São Paulo (UNIFESP) |
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UNIFESP |
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UNIFESP |
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Repositório Institucional da UNIFESP |
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Repositório Institucional da UNIFESP |
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