Characterization of thrombin inhibitory mechanism of rAaTI, a Kazal-type inhibitor from Aedes aegypti with anticoagulant activity
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/11600/33479 http://dx.doi.org/10.1016/j.biochi.2010.12.006 |
Resumo: | Saliva of blood-sucking arthropods contains a complex mixture of anti-haemostatic, anti-inflammatory and immune-modulator compounds. Among anti-haemostatic factors, there are anticoagulants, vasodilators and platelet aggregation inhibitors. Previous analyses of the sialotranscriptome of Aedes aegypti showed the potential presence of a Kazal-type serine protease inhibitor in the female salivary glands, carcass and also in the whole male, which inhibitor we named AaTI (A. aegypti thrombin inhibitor). Recently, we expressed and characterized rAaTI as a trypsin inhibitor, and its anticoagulant activity [1]. in this work we characterized the thrombin inhibition mechanism of rAaTI. Recombinant AaTI was able to prolong prothrombin time, activated partial thromboplastin time and thrombin time. in contrast, AaTI Delta (rAaTI truncated form) and C-terminal AaTI acidic tail prolong only thrombin time. in the competition assay, rAaTI, AaTI Delta or C-terminal AaTI acidic tail thrombin interactions seem to be affected by heparin but not by hirudin, suggesting that rAaTI binds to thrombin exosite 2. Finally, the thrombin inhibition assay of rAaTI showed an uncompetitive inhibition mechanism. in conclusion, rAaTI can probably inhibit thrombin by interacting with thrombin exosite 2, and the interaction is not mediated by the AaTI C-terminal region, since the truncated AaTI Delta form also prolongs thrombin time. (C) 2010 Elsevier Masson SAS. All rights reserved. |
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Watanabe, Renata Midori Okuta [UNIFESP]Tanaka-Azevedo, Anita M.Araujo, Mariana da Silva [UNIFESP]Juliano, Maria Aparecida [UNIFESP]Tanaka, Aparecida Sadae [UNIFESP]Universidade Federal de São Paulo (UNIFESP)Inst Butantan2016-01-24T14:06:12Z2016-01-24T14:06:12Z2011-03-01Biochimie. Paris: Elsevier France-editions Scientifiques Medicales Elsevier, v. 93, n. 3, p. 618-623, 2011.0300-9084http://repositorio.unifesp.br/handle/11600/33479http://dx.doi.org/10.1016/j.biochi.2010.12.006WOS000288405600029.pdf10.1016/j.biochi.2010.12.006WOS:000288405600029Saliva of blood-sucking arthropods contains a complex mixture of anti-haemostatic, anti-inflammatory and immune-modulator compounds. Among anti-haemostatic factors, there are anticoagulants, vasodilators and platelet aggregation inhibitors. Previous analyses of the sialotranscriptome of Aedes aegypti showed the potential presence of a Kazal-type serine protease inhibitor in the female salivary glands, carcass and also in the whole male, which inhibitor we named AaTI (A. aegypti thrombin inhibitor). Recently, we expressed and characterized rAaTI as a trypsin inhibitor, and its anticoagulant activity [1]. in this work we characterized the thrombin inhibition mechanism of rAaTI. Recombinant AaTI was able to prolong prothrombin time, activated partial thromboplastin time and thrombin time. in contrast, AaTI Delta (rAaTI truncated form) and C-terminal AaTI acidic tail prolong only thrombin time. in the competition assay, rAaTI, AaTI Delta or C-terminal AaTI acidic tail thrombin interactions seem to be affected by heparin but not by hirudin, suggesting that rAaTI binds to thrombin exosite 2. Finally, the thrombin inhibition assay of rAaTI showed an uncompetitive inhibition mechanism. in conclusion, rAaTI can probably inhibit thrombin by interacting with thrombin exosite 2, and the interaction is not mediated by the AaTI C-terminal region, since the truncated AaTI Delta form also prolongs thrombin time. (C) 2010 Elsevier Masson SAS. All rights reserved.Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Universidade Federal de São Paulo, Dept Bioquim, BR-04044020 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biofis, BR-04044020 São Paulo, BrazilInst Butantan, Lab Fisiopatol, BR-05503900 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Bioquim, BR-04044020 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biofis, BR-04044020 São Paulo, BrazilFAPESP: 05/03514-9FAPESP: 07/56614-6CNPq: 470070/2004-8CNPq: 575829/2008-7Web of Science618-623engElsevier B.V.Biochimiehttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policyinfo:eu-repo/semantics/openAccessAedes aegyptiKazal-type inhibitorMidgutSalivary glandThrombin inhibitorCharacterization of thrombin inhibitory mechanism of rAaTI, a Kazal-type inhibitor from Aedes aegypti with anticoagulant activityinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlereponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESPORIGINALWOS000288405600029.pdfapplication/pdf611598${dspace.ui.url}/bitstream/11600/33479/1/WOS000288405600029.pdfa2e89d064d64ab048fa5c7fbae4b1162MD51open accessTEXTWOS000288405600029.pdf.txtWOS000288405600029.pdf.txtExtracted texttext/plain30229${dspace.ui.url}/bitstream/11600/33479/2/WOS000288405600029.pdf.txt61e1702af47db3e43ed736d7c1817557MD52open access11600/334792023-02-15 21:48:28.86open accessoai:repositorio.unifesp.br:11600/33479Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestopendoar:34652023-05-25T12:44:50.694816Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false |
dc.title.en.fl_str_mv |
Characterization of thrombin inhibitory mechanism of rAaTI, a Kazal-type inhibitor from Aedes aegypti with anticoagulant activity |
title |
Characterization of thrombin inhibitory mechanism of rAaTI, a Kazal-type inhibitor from Aedes aegypti with anticoagulant activity |
spellingShingle |
Characterization of thrombin inhibitory mechanism of rAaTI, a Kazal-type inhibitor from Aedes aegypti with anticoagulant activity Watanabe, Renata Midori Okuta [UNIFESP] Aedes aegypti Kazal-type inhibitor Midgut Salivary gland Thrombin inhibitor |
title_short |
Characterization of thrombin inhibitory mechanism of rAaTI, a Kazal-type inhibitor from Aedes aegypti with anticoagulant activity |
title_full |
Characterization of thrombin inhibitory mechanism of rAaTI, a Kazal-type inhibitor from Aedes aegypti with anticoagulant activity |
title_fullStr |
Characterization of thrombin inhibitory mechanism of rAaTI, a Kazal-type inhibitor from Aedes aegypti with anticoagulant activity |
title_full_unstemmed |
Characterization of thrombin inhibitory mechanism of rAaTI, a Kazal-type inhibitor from Aedes aegypti with anticoagulant activity |
title_sort |
Characterization of thrombin inhibitory mechanism of rAaTI, a Kazal-type inhibitor from Aedes aegypti with anticoagulant activity |
author |
Watanabe, Renata Midori Okuta [UNIFESP] |
author_facet |
Watanabe, Renata Midori Okuta [UNIFESP] Tanaka-Azevedo, Anita M. Araujo, Mariana da Silva [UNIFESP] Juliano, Maria Aparecida [UNIFESP] Tanaka, Aparecida Sadae [UNIFESP] |
author_role |
author |
author2 |
Tanaka-Azevedo, Anita M. Araujo, Mariana da Silva [UNIFESP] Juliano, Maria Aparecida [UNIFESP] Tanaka, Aparecida Sadae [UNIFESP] |
author2_role |
author author author author |
dc.contributor.institution.none.fl_str_mv |
Universidade Federal de São Paulo (UNIFESP) Inst Butantan |
dc.contributor.author.fl_str_mv |
Watanabe, Renata Midori Okuta [UNIFESP] Tanaka-Azevedo, Anita M. Araujo, Mariana da Silva [UNIFESP] Juliano, Maria Aparecida [UNIFESP] Tanaka, Aparecida Sadae [UNIFESP] |
dc.subject.eng.fl_str_mv |
Aedes aegypti Kazal-type inhibitor Midgut Salivary gland Thrombin inhibitor |
topic |
Aedes aegypti Kazal-type inhibitor Midgut Salivary gland Thrombin inhibitor |
description |
Saliva of blood-sucking arthropods contains a complex mixture of anti-haemostatic, anti-inflammatory and immune-modulator compounds. Among anti-haemostatic factors, there are anticoagulants, vasodilators and platelet aggregation inhibitors. Previous analyses of the sialotranscriptome of Aedes aegypti showed the potential presence of a Kazal-type serine protease inhibitor in the female salivary glands, carcass and also in the whole male, which inhibitor we named AaTI (A. aegypti thrombin inhibitor). Recently, we expressed and characterized rAaTI as a trypsin inhibitor, and its anticoagulant activity [1]. in this work we characterized the thrombin inhibition mechanism of rAaTI. Recombinant AaTI was able to prolong prothrombin time, activated partial thromboplastin time and thrombin time. in contrast, AaTI Delta (rAaTI truncated form) and C-terminal AaTI acidic tail prolong only thrombin time. in the competition assay, rAaTI, AaTI Delta or C-terminal AaTI acidic tail thrombin interactions seem to be affected by heparin but not by hirudin, suggesting that rAaTI binds to thrombin exosite 2. Finally, the thrombin inhibition assay of rAaTI showed an uncompetitive inhibition mechanism. in conclusion, rAaTI can probably inhibit thrombin by interacting with thrombin exosite 2, and the interaction is not mediated by the AaTI C-terminal region, since the truncated AaTI Delta form also prolongs thrombin time. (C) 2010 Elsevier Masson SAS. All rights reserved. |
publishDate |
2011 |
dc.date.issued.fl_str_mv |
2011-03-01 |
dc.date.accessioned.fl_str_mv |
2016-01-24T14:06:12Z |
dc.date.available.fl_str_mv |
2016-01-24T14:06:12Z |
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 |
Biochimie. Paris: Elsevier France-editions Scientifiques Medicales Elsevier, v. 93, n. 3, p. 618-623, 2011. |
dc.identifier.uri.fl_str_mv |
http://repositorio.unifesp.br/handle/11600/33479 http://dx.doi.org/10.1016/j.biochi.2010.12.006 |
dc.identifier.issn.none.fl_str_mv |
0300-9084 |
dc.identifier.file.none.fl_str_mv |
WOS000288405600029.pdf |
dc.identifier.doi.none.fl_str_mv |
10.1016/j.biochi.2010.12.006 |
dc.identifier.wos.none.fl_str_mv |
WOS:000288405600029 |
identifier_str_mv |
Biochimie. Paris: Elsevier France-editions Scientifiques Medicales Elsevier, v. 93, n. 3, p. 618-623, 2011. 0300-9084 WOS000288405600029.pdf 10.1016/j.biochi.2010.12.006 WOS:000288405600029 |
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http://repositorio.unifesp.br/handle/11600/33479 http://dx.doi.org/10.1016/j.biochi.2010.12.006 |
dc.language.iso.fl_str_mv |
eng |
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Biochimie |
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http://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy info:eu-repo/semantics/openAccess |
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http://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy |
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openAccess |
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618-623 |
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Elsevier B.V. |
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Elsevier B.V. |
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