Heparin Modulates the Endopeptidase Activity of Leishmania mexicana Cysteine Protease Cathepsin L-Like rCPB2.8
Autor(a) principal: | |
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Data de Publicação: | 2013 |
Outros Autores: | , , , , , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNIFESP |
Texto Completo: | http://repositorio.unifesp.br/handle/11600/36988 http://dx.doi.org/10.1371/journal.pone.0080153 |
Resumo: | Background: Cysteine protease B is considered crucial for the survival and infectivity of the Leishmania in its human host. Several microorganism pathogens bind to the heparin-like glycosaminoglycans chains of proteoglycans at host-cell surface to promote their attachment and internalization. Here, we have investigated the influence of heparin upon Leishmania mexicana cysteine protease rCPB2.8 activity.Methodology/Principal Findings: the data analysis revealed that the presence of heparin affects all steps of the enzyme reaction: (i) it decreases 3.5-fold the k(1) and 4.0-fold the k(-1), (ii) it affects the acyl-enzyme accumulation with pronounced decrease in k(2) (2.7-fold), and also decrease in k(3) (3.5-fold). the large values of triangle G = 12 kJ/mol for the association and dissociation steps indicate substantial structural strains linked to the formation/dissociation of the ES complex in the presence of heparin, which underscore a conformational change that prevents the diffusion of substrate in the rCPB2.8 active site. Binding to heparin also significantly decreases the alpha-helix content of the rCPB2.8 and perturbs the intrinsic fluorescence emission of the enzyme. the data strongly suggest that heparin is altering the ionization of catalytic (Cys(25))-S-/(His(163))-Im(+) H ion pair of the rCPB2.8. Moreover, the interaction of heparin with the N-terminal pro-region of rCPB2.8 significantly decreased its inhibitory activity against the mature enzyme.Conclusions/Significance: Taken together, depending on their concentration, heparin-like glycosaminoglycans can either stimulate or antagonize the activity of cysteine protease B enzymes during parasite infection, suggesting that this glycoconjugate can anchor parasite cysteine protease at host cell surface. |
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Judice, Wagner A. S.Manfredi, Marcella A.Souza, Gerson P.Sansevero, Thiago M.Almeida, Paulo C. [UNIFESP]Shida, Claudio S. [UNIFESP]Gesteira, Tarsis F.Juliano, Luiz [UNIFESP]Westrop, Gareth D.Sanderson, Sanya J.Coombs, Graham H.Tersariol, Ivarne Luis dos Santos [UNIFESP]Univ Mogi das CruzesUniversidade Federal de São Paulo (UNIFESP)Cincinnati Childrens Hosp Med CtrUniv Strathclyde2016-01-24T14:34:43Z2016-01-24T14:34:43Z2013-11-21Plos One. San Francisco: Public Library Science, v. 8, n. 11, 12 p., 2013.1932-6203http://repositorio.unifesp.br/handle/11600/36988http://dx.doi.org/10.1371/journal.pone.0080153WOS000327539800059.pdf10.1371/journal.pone.0080153WOS:000327539800059Background: Cysteine protease B is considered crucial for the survival and infectivity of the Leishmania in its human host. Several microorganism pathogens bind to the heparin-like glycosaminoglycans chains of proteoglycans at host-cell surface to promote their attachment and internalization. Here, we have investigated the influence of heparin upon Leishmania mexicana cysteine protease rCPB2.8 activity.Methodology/Principal Findings: the data analysis revealed that the presence of heparin affects all steps of the enzyme reaction: (i) it decreases 3.5-fold the k(1) and 4.0-fold the k(-1), (ii) it affects the acyl-enzyme accumulation with pronounced decrease in k(2) (2.7-fold), and also decrease in k(3) (3.5-fold). the large values of triangle G = 12 kJ/mol for the association and dissociation steps indicate substantial structural strains linked to the formation/dissociation of the ES complex in the presence of heparin, which underscore a conformational change that prevents the diffusion of substrate in the rCPB2.8 active site. Binding to heparin also significantly decreases the alpha-helix content of the rCPB2.8 and perturbs the intrinsic fluorescence emission of the enzyme. the data strongly suggest that heparin is altering the ionization of catalytic (Cys(25))-S-/(His(163))-Im(+) H ion pair of the rCPB2.8. Moreover, the interaction of heparin with the N-terminal pro-region of rCPB2.8 significantly decreased its inhibitory activity against the mature enzyme.Conclusions/Significance: Taken together, depending on their concentration, heparin-like glycosaminoglycans can either stimulate or antagonize the activity of cysteine protease B enzymes during parasite infection, suggesting that this glycoconjugate can anchor parasite cysteine protease at host cell surface.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Cientifico TecnologicoMedical Research CouncilUniv Mogi das Cruzes, Ctr Interdisciplinar Invest Bioquim, Mogi Das Cruzes, BrazilUniversidade Federal de São Paulo, Dept Bioquim, São Paulo, BrazilUniversidade Federal de São Paulo, Inst Ciencia & Tecnol, Sao Jose Dos Campos, BrazilCincinnati Childrens Hosp Med Ctr, Div Dev Biol, Cincinnati, OH 45229 USAUniversidade Federal de São Paulo, Dept Biofis, São Paulo, BrazilUniv Strathclyde, Strathclyde Inst Pharm & Biomed Sci, Glasgow, Lanark, ScotlandUniversidade Federal de São Paulo, Dept Bioquim, São Paulo, BrazilUniversidade Federal de São Paulo, Inst Ciencia & Tecnol, Sao Jose Dos Campos, BrazilUniversidade Federal de São Paulo, Dept Biofis, São Paulo, BrazilFAPESP: 2012/50219-6Conselho Nacional de Desenvolvimento Cientifico Tecnologico: 303843/2009-8Medical Research Council: G0700127Web of Science12engPublic Library SciencePlos OneHeparin Modulates the Endopeptidase Activity of Leishmania mexicana Cysteine Protease Cathepsin L-Like rCPB2.8info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESPORIGINALWOS000327539800059.pdfapplication/pdf1535588${dspace.ui.url}/bitstream/11600/36988/1/WOS000327539800059.pdfb1cdcf7822973d8648b083bd6b90803dMD51open accessTEXTWOS000327539800059.pdf.txtWOS000327539800059.pdf.txtExtracted texttext/plain70319${dspace.ui.url}/bitstream/11600/36988/9/WOS000327539800059.pdf.txte1e1744f7846e5cd99839c834bc6a6ceMD59open accessTHUMBNAILWOS000327539800059.pdf.jpgWOS000327539800059.pdf.jpgIM Thumbnailimage/jpeg7960${dspace.ui.url}/bitstream/11600/36988/11/WOS000327539800059.pdf.jpgb82c8b548c60d2f197486403ebc7fca2MD511open access11600/369882023-06-05 19:05:47.708open accessoai:repositorio.unifesp.br:11600/36988Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestopendoar:34652023-06-05T22:05:47Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false |
dc.title.en.fl_str_mv |
Heparin Modulates the Endopeptidase Activity of Leishmania mexicana Cysteine Protease Cathepsin L-Like rCPB2.8 |
title |
Heparin Modulates the Endopeptidase Activity of Leishmania mexicana Cysteine Protease Cathepsin L-Like rCPB2.8 |
spellingShingle |
Heparin Modulates the Endopeptidase Activity of Leishmania mexicana Cysteine Protease Cathepsin L-Like rCPB2.8 Judice, Wagner A. S. |
title_short |
Heparin Modulates the Endopeptidase Activity of Leishmania mexicana Cysteine Protease Cathepsin L-Like rCPB2.8 |
title_full |
Heparin Modulates the Endopeptidase Activity of Leishmania mexicana Cysteine Protease Cathepsin L-Like rCPB2.8 |
title_fullStr |
Heparin Modulates the Endopeptidase Activity of Leishmania mexicana Cysteine Protease Cathepsin L-Like rCPB2.8 |
title_full_unstemmed |
Heparin Modulates the Endopeptidase Activity of Leishmania mexicana Cysteine Protease Cathepsin L-Like rCPB2.8 |
title_sort |
Heparin Modulates the Endopeptidase Activity of Leishmania mexicana Cysteine Protease Cathepsin L-Like rCPB2.8 |
author |
Judice, Wagner A. S. |
author_facet |
Judice, Wagner A. S. Manfredi, Marcella A. Souza, Gerson P. Sansevero, Thiago M. Almeida, Paulo C. [UNIFESP] Shida, Claudio S. [UNIFESP] Gesteira, Tarsis F. Juliano, Luiz [UNIFESP] Westrop, Gareth D. Sanderson, Sanya J. Coombs, Graham H. Tersariol, Ivarne Luis dos Santos [UNIFESP] |
author_role |
author |
author2 |
Manfredi, Marcella A. Souza, Gerson P. Sansevero, Thiago M. Almeida, Paulo C. [UNIFESP] Shida, Claudio S. [UNIFESP] Gesteira, Tarsis F. Juliano, Luiz [UNIFESP] Westrop, Gareth D. Sanderson, Sanya J. Coombs, Graham H. Tersariol, Ivarne Luis dos Santos [UNIFESP] |
author2_role |
author author author author author author author author author author author |
dc.contributor.institution.none.fl_str_mv |
Univ Mogi das Cruzes Universidade Federal de São Paulo (UNIFESP) Cincinnati Childrens Hosp Med Ctr Univ Strathclyde |
dc.contributor.author.fl_str_mv |
Judice, Wagner A. S. Manfredi, Marcella A. Souza, Gerson P. Sansevero, Thiago M. Almeida, Paulo C. [UNIFESP] Shida, Claudio S. [UNIFESP] Gesteira, Tarsis F. Juliano, Luiz [UNIFESP] Westrop, Gareth D. Sanderson, Sanya J. Coombs, Graham H. Tersariol, Ivarne Luis dos Santos [UNIFESP] |
description |
Background: Cysteine protease B is considered crucial for the survival and infectivity of the Leishmania in its human host. Several microorganism pathogens bind to the heparin-like glycosaminoglycans chains of proteoglycans at host-cell surface to promote their attachment and internalization. Here, we have investigated the influence of heparin upon Leishmania mexicana cysteine protease rCPB2.8 activity.Methodology/Principal Findings: the data analysis revealed that the presence of heparin affects all steps of the enzyme reaction: (i) it decreases 3.5-fold the k(1) and 4.0-fold the k(-1), (ii) it affects the acyl-enzyme accumulation with pronounced decrease in k(2) (2.7-fold), and also decrease in k(3) (3.5-fold). the large values of triangle G = 12 kJ/mol for the association and dissociation steps indicate substantial structural strains linked to the formation/dissociation of the ES complex in the presence of heparin, which underscore a conformational change that prevents the diffusion of substrate in the rCPB2.8 active site. Binding to heparin also significantly decreases the alpha-helix content of the rCPB2.8 and perturbs the intrinsic fluorescence emission of the enzyme. the data strongly suggest that heparin is altering the ionization of catalytic (Cys(25))-S-/(His(163))-Im(+) H ion pair of the rCPB2.8. Moreover, the interaction of heparin with the N-terminal pro-region of rCPB2.8 significantly decreased its inhibitory activity against the mature enzyme.Conclusions/Significance: Taken together, depending on their concentration, heparin-like glycosaminoglycans can either stimulate or antagonize the activity of cysteine protease B enzymes during parasite infection, suggesting that this glycoconjugate can anchor parasite cysteine protease at host cell surface. |
publishDate |
2013 |
dc.date.issued.fl_str_mv |
2013-11-21 |
dc.date.accessioned.fl_str_mv |
2016-01-24T14:34:43Z |
dc.date.available.fl_str_mv |
2016-01-24T14:34:43Z |
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info:eu-repo/semantics/publishedVersion |
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format |
article |
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dc.identifier.citation.fl_str_mv |
Plos One. San Francisco: Public Library Science, v. 8, n. 11, 12 p., 2013. |
dc.identifier.uri.fl_str_mv |
http://repositorio.unifesp.br/handle/11600/36988 http://dx.doi.org/10.1371/journal.pone.0080153 |
dc.identifier.issn.none.fl_str_mv |
1932-6203 |
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WOS000327539800059.pdf |
dc.identifier.doi.none.fl_str_mv |
10.1371/journal.pone.0080153 |
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WOS:000327539800059 |
identifier_str_mv |
Plos One. San Francisco: Public Library Science, v. 8, n. 11, 12 p., 2013. 1932-6203 WOS000327539800059.pdf 10.1371/journal.pone.0080153 WOS:000327539800059 |
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http://repositorio.unifesp.br/handle/11600/36988 http://dx.doi.org/10.1371/journal.pone.0080153 |
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Public Library Science |
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Public Library Science |
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