Identification and characterization of excretory/secretory products of Giardia duodenalis trophozoites
Autor(a) principal: | |
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Data de Publicação: | 2008 |
Tipo de documento: | Artigo de conferência |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1590/S1678-91992008000300016 http://hdl.handle.net/11449/18944 |
Resumo: | One of the major questions concerning Giardia is the understanding of pathophysiological processes associated with small intestine abnormalities. There are evidences that Giardia trophozoites contain and/or release proteolytic enzymes that may be implicated in the host intestinal epithelium. The present investigation was undertaken to examine the protease activity in excretory/secretory (E/S) products of Giardia duodenalis trophozoites of an axenic Brazilian strain (BTU-11) and the reference strain Portland 1 (P1). E/S products from trophozoites of each strain in conditioned medium were tested with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) for the protein profiles, and the protease activity was analyzed using substrate-impregnated SDS-PAGE (gelatin and collagen) and hemoglobin assay. The proteases characterization was based on inhibition assays including synthetic inhibitors. Electrophoresis analysis of E/S products revealed a banding pattern composed by few bands (4 to 6 bands) in the migration region of 123 to 28 kDa. Proteolytic products were detected in the conditioned medium by trophozoites of both assayed strains. In the gels containing copolymerized gelatin and collagen, E/S products promoted substrate degradation and the most evident proteolysis zones were distributed in the migration regions of 77 to 18 kDa and 145 to 18 kDa, respectively, in the patterns of gelatinolytic and collagenolytic activities. Degradation of hemoglobin was also observed, and the pattern of hydrolysis was similar in both E/S products assayed. Inhibitor assays showed that the main proteolytic activity in both E/S products is due to cysteine proteases, although the presence of serine proteases was also indicated. Degradation of substrates including collagen and hemoglobin could lead us to speculate different functions of Giardia excreted/secreted proteases in vivo, but to confirm this possibility and to elucidate its implication on host-parasite interactions, further experiments applying protocols for the purification of proteases are necessary. Even so, our observations are relevant and hold the perspective for the understanding about protease activity in Giardia trophozoites of axenic strain isolated in an endemic area. |
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Identification and characterization of excretory/secretory products of Giardia duodenalis trophozoitesGiardia duodenalistrophozoitesexcretory/secretory productsproteasesprotein substratesOne of the major questions concerning Giardia is the understanding of pathophysiological processes associated with small intestine abnormalities. There are evidences that Giardia trophozoites contain and/or release proteolytic enzymes that may be implicated in the host intestinal epithelium. The present investigation was undertaken to examine the protease activity in excretory/secretory (E/S) products of Giardia duodenalis trophozoites of an axenic Brazilian strain (BTU-11) and the reference strain Portland 1 (P1). E/S products from trophozoites of each strain in conditioned medium were tested with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) for the protein profiles, and the protease activity was analyzed using substrate-impregnated SDS-PAGE (gelatin and collagen) and hemoglobin assay. The proteases characterization was based on inhibition assays including synthetic inhibitors. Electrophoresis analysis of E/S products revealed a banding pattern composed by few bands (4 to 6 bands) in the migration region of 123 to 28 kDa. Proteolytic products were detected in the conditioned medium by trophozoites of both assayed strains. In the gels containing copolymerized gelatin and collagen, E/S products promoted substrate degradation and the most evident proteolysis zones were distributed in the migration regions of 77 to 18 kDa and 145 to 18 kDa, respectively, in the patterns of gelatinolytic and collagenolytic activities. Degradation of hemoglobin was also observed, and the pattern of hydrolysis was similar in both E/S products assayed. Inhibitor assays showed that the main proteolytic activity in both E/S products is due to cysteine proteases, although the presence of serine proteases was also indicated. Degradation of substrates including collagen and hemoglobin could lead us to speculate different functions of Giardia excreted/secreted proteases in vivo, but to confirm this possibility and to elucidate its implication on host-parasite interactions, further experiments applying protocols for the purification of proteases are necessary. Even so, our observations are relevant and hold the perspective for the understanding about protease activity in Giardia trophozoites of axenic strain isolated in an endemic area.UNESP, Inst Biociencias, Dept Parasitol, BR-18618000 Botucatu, SP, BrazilUNESP, Inst Biociencias, Dept Parasitol, BR-18618000 Botucatu, SP, BrazilUniversidade Estadual Paulista (Unesp), Centro de Estudos de Venenos e Animais Peçonhentos (CEVAP)Universidade Estadual Paulista (Unesp)De Carvalho, Thais Batista [UNESP]2014-05-20T13:53:06Z2014-05-20T13:53:06Z2008-01-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObject553-U6application/pdfhttp://dx.doi.org/10.1590/S1678-91992008000300016Journal of Venomous Animals and Toxins Including Tropical Diseases. Botucatu: Cevap-unesp, v. 14, n. 3, p. 553-U6, 2008.1678-9199http://hdl.handle.net/11449/18944S1678-91992008000300016WOS:000259279500016S1678-91992008000300016.pdfWeb of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengJournal of Venomous Animals and Toxins Including Tropical Diseases1.7820,573info:eu-repo/semantics/openAccess2024-01-08T06:26:49Zoai:repositorio.unesp.br:11449/18944Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T22:26:56.711549Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Identification and characterization of excretory/secretory products of Giardia duodenalis trophozoites |
title |
Identification and characterization of excretory/secretory products of Giardia duodenalis trophozoites |
spellingShingle |
Identification and characterization of excretory/secretory products of Giardia duodenalis trophozoites De Carvalho, Thais Batista [UNESP] Giardia duodenalis trophozoites excretory/secretory products proteases protein substrates |
title_short |
Identification and characterization of excretory/secretory products of Giardia duodenalis trophozoites |
title_full |
Identification and characterization of excretory/secretory products of Giardia duodenalis trophozoites |
title_fullStr |
Identification and characterization of excretory/secretory products of Giardia duodenalis trophozoites |
title_full_unstemmed |
Identification and characterization of excretory/secretory products of Giardia duodenalis trophozoites |
title_sort |
Identification and characterization of excretory/secretory products of Giardia duodenalis trophozoites |
author |
De Carvalho, Thais Batista [UNESP] |
author_facet |
De Carvalho, Thais Batista [UNESP] |
author_role |
author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (Unesp) |
dc.contributor.author.fl_str_mv |
De Carvalho, Thais Batista [UNESP] |
dc.subject.por.fl_str_mv |
Giardia duodenalis trophozoites excretory/secretory products proteases protein substrates |
topic |
Giardia duodenalis trophozoites excretory/secretory products proteases protein substrates |
description |
One of the major questions concerning Giardia is the understanding of pathophysiological processes associated with small intestine abnormalities. There are evidences that Giardia trophozoites contain and/or release proteolytic enzymes that may be implicated in the host intestinal epithelium. The present investigation was undertaken to examine the protease activity in excretory/secretory (E/S) products of Giardia duodenalis trophozoites of an axenic Brazilian strain (BTU-11) and the reference strain Portland 1 (P1). E/S products from trophozoites of each strain in conditioned medium were tested with sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) for the protein profiles, and the protease activity was analyzed using substrate-impregnated SDS-PAGE (gelatin and collagen) and hemoglobin assay. The proteases characterization was based on inhibition assays including synthetic inhibitors. Electrophoresis analysis of E/S products revealed a banding pattern composed by few bands (4 to 6 bands) in the migration region of 123 to 28 kDa. Proteolytic products were detected in the conditioned medium by trophozoites of both assayed strains. In the gels containing copolymerized gelatin and collagen, E/S products promoted substrate degradation and the most evident proteolysis zones were distributed in the migration regions of 77 to 18 kDa and 145 to 18 kDa, respectively, in the patterns of gelatinolytic and collagenolytic activities. Degradation of hemoglobin was also observed, and the pattern of hydrolysis was similar in both E/S products assayed. Inhibitor assays showed that the main proteolytic activity in both E/S products is due to cysteine proteases, although the presence of serine proteases was also indicated. Degradation of substrates including collagen and hemoglobin could lead us to speculate different functions of Giardia excreted/secreted proteases in vivo, but to confirm this possibility and to elucidate its implication on host-parasite interactions, further experiments applying protocols for the purification of proteases are necessary. Even so, our observations are relevant and hold the perspective for the understanding about protease activity in Giardia trophozoites of axenic strain isolated in an endemic area. |
publishDate |
2008 |
dc.date.none.fl_str_mv |
2008-01-01 2014-05-20T13:53:06Z 2014-05-20T13:53:06Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/conferenceObject |
format |
conferenceObject |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://dx.doi.org/10.1590/S1678-91992008000300016 Journal of Venomous Animals and Toxins Including Tropical Diseases. Botucatu: Cevap-unesp, v. 14, n. 3, p. 553-U6, 2008. 1678-9199 http://hdl.handle.net/11449/18944 S1678-91992008000300016 WOS:000259279500016 S1678-91992008000300016.pdf |
url |
http://dx.doi.org/10.1590/S1678-91992008000300016 http://hdl.handle.net/11449/18944 |
identifier_str_mv |
Journal of Venomous Animals and Toxins Including Tropical Diseases. Botucatu: Cevap-unesp, v. 14, n. 3, p. 553-U6, 2008. 1678-9199 S1678-91992008000300016 WOS:000259279500016 S1678-91992008000300016.pdf |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Journal of Venomous Animals and Toxins Including Tropical Diseases 1.782 0,573 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
553-U6 application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Estadual Paulista (Unesp), Centro de Estudos de Venenos e Animais Peçonhentos (CEVAP) |
publisher.none.fl_str_mv |
Universidade Estadual Paulista (Unesp), Centro de Estudos de Venenos e Animais Peçonhentos (CEVAP) |
dc.source.none.fl_str_mv |
Web of Science reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
|
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1808129428053557248 |