Proteolytic activity in the adult and larval stages of the human roundworm parasite Angiostrongylus costaricensis
Autor(a) principal: | |
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Data de Publicação: | 2012 |
Outros Autores: | , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Memórias do Instituto Oswaldo Cruz |
Texto Completo: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762012000600008 |
Resumo: | Angiostrongylus costaricensis is a nematode that causes abdominal angiostrongyliasis, a widespread human parasitism in Latin America. This study aimed to characterize the protease profiles of different developmental stages of this helminth. First-stage larvae (L1) were obtained from the faeces of infected Sigmodon hispidus rodents and third-stage larvae (L3) were collected from mollusks Biomphalaria glabrata previously infected with L1. Adult worms were recovered from rodent mesenteric arteries. Protein extraction was performed after repeated freeze-thaw cycles followed by maceration of the nematodes in 40 mM Tris base. Proteolysis of gelatin was observed by zymography and found only in the larval stages. In L3, the gelatinolytic activity was effectively inhibited by orthophenanthroline, indicating the involvement of metalloproteases. The mechanistic class of the gelatinases from L1 could not be precisely determined using traditional class-specific inhibitors. Adult worm extracts were able to hydrolyze haemoglobin in solution, although no activity was observed by zymography. This haemoglobinolytic activity was ascribed to aspartic proteases following its effective inhibition by pepstatin, which also inhibited the haemoglobinolytic activity of L1 and L3 extracts. The characterization of protease expression throughout the A. costaricensis life cycle may reveal key factors influencing the process of parasitic infection and thus foster our understanding of the disease pathogenesis. |
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Memórias do Instituto Oswaldo Cruz |
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Proteolytic activity in the adult and larval stages of the human roundworm parasite Angiostrongylus costaricensisAngiostrongylus costaricensismetalloproteasesaspartic proteaseshaemoglobinzymographyAngiostrongylus costaricensis is a nematode that causes abdominal angiostrongyliasis, a widespread human parasitism in Latin America. This study aimed to characterize the protease profiles of different developmental stages of this helminth. First-stage larvae (L1) were obtained from the faeces of infected Sigmodon hispidus rodents and third-stage larvae (L3) were collected from mollusks Biomphalaria glabrata previously infected with L1. Adult worms were recovered from rodent mesenteric arteries. Protein extraction was performed after repeated freeze-thaw cycles followed by maceration of the nematodes in 40 mM Tris base. Proteolysis of gelatin was observed by zymography and found only in the larval stages. In L3, the gelatinolytic activity was effectively inhibited by orthophenanthroline, indicating the involvement of metalloproteases. The mechanistic class of the gelatinases from L1 could not be precisely determined using traditional class-specific inhibitors. Adult worm extracts were able to hydrolyze haemoglobin in solution, although no activity was observed by zymography. This haemoglobinolytic activity was ascribed to aspartic proteases following its effective inhibition by pepstatin, which also inhibited the haemoglobinolytic activity of L1 and L3 extracts. The characterization of protease expression throughout the A. costaricensis life cycle may reveal key factors influencing the process of parasitic infection and thus foster our understanding of the disease pathogenesis.Instituto Oswaldo Cruz, Ministério da Saúde2012-09-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersiontext/htmlhttp://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762012000600008Memórias do Instituto Oswaldo Cruz v.107 n.6 2012reponame:Memórias do Instituto Oswaldo Cruzinstname:Fundação Oswaldo Cruzinstacron:FIOCRUZ10.1590/S0074-02762012000600008info:eu-repo/semantics/openAccessRebello,Karina MastropasquaSiqueira,Caroline Reis deRibeiro,Erika LouiseValente,Richard HemmiMota,Ester MariaPerales,JonasNeves-Ferreira,Ana Gisele da CostaLenzi,Henrique Leoneleng2020-04-25T17:51:15Zhttp://www.scielo.br/oai/scielo-oai.php0074-02761678-8060opendoar:null2020-04-26 02:18:29.716Memórias do Instituto Oswaldo Cruz - Fundação Oswaldo Cruztrue |
dc.title.none.fl_str_mv |
Proteolytic activity in the adult and larval stages of the human roundworm parasite Angiostrongylus costaricensis |
title |
Proteolytic activity in the adult and larval stages of the human roundworm parasite Angiostrongylus costaricensis |
spellingShingle |
Proteolytic activity in the adult and larval stages of the human roundworm parasite Angiostrongylus costaricensis Rebello,Karina Mastropasqua Angiostrongylus costaricensis metalloproteases aspartic proteases haemoglobin zymography |
title_short |
Proteolytic activity in the adult and larval stages of the human roundworm parasite Angiostrongylus costaricensis |
title_full |
Proteolytic activity in the adult and larval stages of the human roundworm parasite Angiostrongylus costaricensis |
title_fullStr |
Proteolytic activity in the adult and larval stages of the human roundworm parasite Angiostrongylus costaricensis |
title_full_unstemmed |
Proteolytic activity in the adult and larval stages of the human roundworm parasite Angiostrongylus costaricensis |
title_sort |
Proteolytic activity in the adult and larval stages of the human roundworm parasite Angiostrongylus costaricensis |
author |
Rebello,Karina Mastropasqua |
author_facet |
Rebello,Karina Mastropasqua Siqueira,Caroline Reis de Ribeiro,Erika Louise Valente,Richard Hemmi Mota,Ester Maria Perales,Jonas Neves-Ferreira,Ana Gisele da Costa Lenzi,Henrique Leonel |
author_role |
author |
author2 |
Siqueira,Caroline Reis de Ribeiro,Erika Louise Valente,Richard Hemmi Mota,Ester Maria Perales,Jonas Neves-Ferreira,Ana Gisele da Costa Lenzi,Henrique Leonel |
author2_role |
author author author author author author author |
dc.contributor.author.fl_str_mv |
Rebello,Karina Mastropasqua Siqueira,Caroline Reis de Ribeiro,Erika Louise Valente,Richard Hemmi Mota,Ester Maria Perales,Jonas Neves-Ferreira,Ana Gisele da Costa Lenzi,Henrique Leonel |
dc.subject.por.fl_str_mv |
Angiostrongylus costaricensis metalloproteases aspartic proteases haemoglobin zymography |
topic |
Angiostrongylus costaricensis metalloproteases aspartic proteases haemoglobin zymography |
dc.description.none.fl_txt_mv |
Angiostrongylus costaricensis is a nematode that causes abdominal angiostrongyliasis, a widespread human parasitism in Latin America. This study aimed to characterize the protease profiles of different developmental stages of this helminth. First-stage larvae (L1) were obtained from the faeces of infected Sigmodon hispidus rodents and third-stage larvae (L3) were collected from mollusks Biomphalaria glabrata previously infected with L1. Adult worms were recovered from rodent mesenteric arteries. Protein extraction was performed after repeated freeze-thaw cycles followed by maceration of the nematodes in 40 mM Tris base. Proteolysis of gelatin was observed by zymography and found only in the larval stages. In L3, the gelatinolytic activity was effectively inhibited by orthophenanthroline, indicating the involvement of metalloproteases. The mechanistic class of the gelatinases from L1 could not be precisely determined using traditional class-specific inhibitors. Adult worm extracts were able to hydrolyze haemoglobin in solution, although no activity was observed by zymography. This haemoglobinolytic activity was ascribed to aspartic proteases following its effective inhibition by pepstatin, which also inhibited the haemoglobinolytic activity of L1 and L3 extracts. The characterization of protease expression throughout the A. costaricensis life cycle may reveal key factors influencing the process of parasitic infection and thus foster our understanding of the disease pathogenesis. |
description |
Angiostrongylus costaricensis is a nematode that causes abdominal angiostrongyliasis, a widespread human parasitism in Latin America. This study aimed to characterize the protease profiles of different developmental stages of this helminth. First-stage larvae (L1) were obtained from the faeces of infected Sigmodon hispidus rodents and third-stage larvae (L3) were collected from mollusks Biomphalaria glabrata previously infected with L1. Adult worms were recovered from rodent mesenteric arteries. Protein extraction was performed after repeated freeze-thaw cycles followed by maceration of the nematodes in 40 mM Tris base. Proteolysis of gelatin was observed by zymography and found only in the larval stages. In L3, the gelatinolytic activity was effectively inhibited by orthophenanthroline, indicating the involvement of metalloproteases. The mechanistic class of the gelatinases from L1 could not be precisely determined using traditional class-specific inhibitors. Adult worm extracts were able to hydrolyze haemoglobin in solution, although no activity was observed by zymography. This haemoglobinolytic activity was ascribed to aspartic proteases following its effective inhibition by pepstatin, which also inhibited the haemoglobinolytic activity of L1 and L3 extracts. The characterization of protease expression throughout the A. costaricensis life cycle may reveal key factors influencing the process of parasitic infection and thus foster our understanding of the disease pathogenesis. |
publishDate |
2012 |
dc.date.none.fl_str_mv |
2012-09-01 |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762012000600008 |
url |
http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0074-02762012000600008 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
10.1590/S0074-02762012000600008 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
text/html |
dc.publisher.none.fl_str_mv |
Instituto Oswaldo Cruz, Ministério da Saúde |
publisher.none.fl_str_mv |
Instituto Oswaldo Cruz, Ministério da Saúde |
dc.source.none.fl_str_mv |
Memórias do Instituto Oswaldo Cruz v.107 n.6 2012 reponame:Memórias do Instituto Oswaldo Cruz instname:Fundação Oswaldo Cruz instacron:FIOCRUZ |
reponame_str |
Memórias do Instituto Oswaldo Cruz |
collection |
Memórias do Instituto Oswaldo Cruz |
instname_str |
Fundação Oswaldo Cruz |
instacron_str |
FIOCRUZ |
institution |
FIOCRUZ |
repository.name.fl_str_mv |
Memórias do Instituto Oswaldo Cruz - Fundação Oswaldo Cruz |
repository.mail.fl_str_mv |
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1669937712126230528 |