Proregion of Acanthoscelides obtectus cysteine proteinase: a novel peptide with enhanced selectivity toward endogenous enzymes
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Publication Date: | 2007 |
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Format: | Article |
Language: | eng |
Source: | Repositório Institucional da UCB |
Download full: | http://twingo.ucb.br:8080/jspui/handle/10869/559 https://repositorio.ucb.br:9443/jspui/handle/123456789/7790 |
Summary: | Acanthoscelides obtectus is a devastating storage insect pest capable of causing severe bean crop losses. In order to maintain their own development, insect pest larvae feed continuously, synthesizing efficient digestive enzymes. Among them, cysteine proteinases (CPs) are commonly produced as inactive precursors (procysteines), requiring a cleavage of the peptide proregion to become active. The proregion fits tightly into the active site of procysteines, efficiently preventing their activity. In this report, a CP cDNA (cpao) was isolated from A. obtectus midgut larvae. In silico studies indicated that the complete CP sequence contains a hydrophobic signal peptide, a prodomain and a conserved catalytic region. Moreover, the encoding cDNA contains 963 bp translating into a 321 residue protein, CPAo, which was expressed in E. coli, fused with thioredoxin. Enzymatic assays using the recombinant protein revealed that the enzyme was catalytically active, being able to cleave the synthetic substrate Z-Phe-Arg-7-AMC. Additionally, this report also focuses the cpao propeptide (PCPAo) subcloning and expression. The expressed propeptide efficiently inhibited CPAo, as well as digestive CP of other bean bruchids. Little or no activity was found against proteolytic enzymes of two other coleopterans: Rhyzopertha dominica and Anthonomus grandis. The data reported here indicate the possibility of endogenous propeptides as a novel strategy on bruchids control, which could be applicable to bean improvement programs. |
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Silva, Francine BarbosaMonteiro, Ana Carolina dos SantosDel Sarto, Rafael PerseghiniMarra, Brener MagnaboscoDias, Simoni C.Figueira, Edson Luiz ZangrandoOliveira, Gustavo Ramos deRocha, Thales LimaSouza, Djair dos Santos deSilva, Maria Cristina Mattar daFranco, Octavio LuizGrossi de Sa, Maria Fatima2016-10-10T03:52:40Z2016-10-10T03:52:40Z2007SILVA, F. B. et al. Proregion of Acanthoscelides obtectus cysteine proteinase: a novel peptide with enhanced selectivity toward endogenous enzymes. Peptidies, v. 28, n. 6, p. 1292-1298, 2007.http://twingo.ucb.br:8080/jspui/handle/10869/559https://repositorio.ucb.br:9443/jspui/handle/123456789/7790Acanthoscelides obtectus is a devastating storage insect pest capable of causing severe bean crop losses. In order to maintain their own development, insect pest larvae feed continuously, synthesizing efficient digestive enzymes. Among them, cysteine proteinases (CPs) are commonly produced as inactive precursors (procysteines), requiring a cleavage of the peptide proregion to become active. The proregion fits tightly into the active site of procysteines, efficiently preventing their activity. In this report, a CP cDNA (cpao) was isolated from A. obtectus midgut larvae. In silico studies indicated that the complete CP sequence contains a hydrophobic signal peptide, a prodomain and a conserved catalytic region. Moreover, the encoding cDNA contains 963 bp translating into a 321 residue protein, CPAo, which was expressed in E. coli, fused with thioredoxin. Enzymatic assays using the recombinant protein revealed that the enzyme was catalytically active, being able to cleave the synthetic substrate Z-Phe-Arg-7-AMC. Additionally, this report also focuses the cpao propeptide (PCPAo) subcloning and expression. The expressed propeptide efficiently inhibited CPAo, as well as digestive CP of other bean bruchids. Little or no activity was found against proteolytic enzymes of two other coleopterans: Rhyzopertha dominica and Anthonomus grandis. The data reported here indicate the possibility of endogenous propeptides as a novel strategy on bruchids control, which could be applicable to bean improvement programs.Made available in DSpace on 2016-10-10T03:52:40Z (GMT). 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dc.title.pt_BR.fl_str_mv |
Proregion of Acanthoscelides obtectus cysteine proteinase: a novel peptide with enhanced selectivity toward endogenous enzymes |
title |
Proregion of Acanthoscelides obtectus cysteine proteinase: a novel peptide with enhanced selectivity toward endogenous enzymes |
spellingShingle |
Proregion of Acanthoscelides obtectus cysteine proteinase: a novel peptide with enhanced selectivity toward endogenous enzymes Silva, Francine Barbosa Acanthoscelides obtectus Cysteine proteinase Propeptide Plant defense |
title_short |
Proregion of Acanthoscelides obtectus cysteine proteinase: a novel peptide with enhanced selectivity toward endogenous enzymes |
title_full |
Proregion of Acanthoscelides obtectus cysteine proteinase: a novel peptide with enhanced selectivity toward endogenous enzymes |
title_fullStr |
Proregion of Acanthoscelides obtectus cysteine proteinase: a novel peptide with enhanced selectivity toward endogenous enzymes |
title_full_unstemmed |
Proregion of Acanthoscelides obtectus cysteine proteinase: a novel peptide with enhanced selectivity toward endogenous enzymes |
title_sort |
Proregion of Acanthoscelides obtectus cysteine proteinase: a novel peptide with enhanced selectivity toward endogenous enzymes |
author |
Silva, Francine Barbosa |
author_facet |
Silva, Francine Barbosa Monteiro, Ana Carolina dos Santos Del Sarto, Rafael Perseghini Marra, Brener Magnabosco Dias, Simoni C. Figueira, Edson Luiz Zangrando Oliveira, Gustavo Ramos de Rocha, Thales Lima Souza, Djair dos Santos de Silva, Maria Cristina Mattar da Franco, Octavio Luiz Grossi de Sa, Maria Fatima |
author_role |
author |
author2 |
Monteiro, Ana Carolina dos Santos Del Sarto, Rafael Perseghini Marra, Brener Magnabosco Dias, Simoni C. Figueira, Edson Luiz Zangrando Oliveira, Gustavo Ramos de Rocha, Thales Lima Souza, Djair dos Santos de Silva, Maria Cristina Mattar da Franco, Octavio Luiz Grossi de Sa, Maria Fatima |
author2_role |
author author author author author author author author author author author |
dc.contributor.author.fl_str_mv |
Silva, Francine Barbosa Monteiro, Ana Carolina dos Santos Del Sarto, Rafael Perseghini Marra, Brener Magnabosco Dias, Simoni C. Figueira, Edson Luiz Zangrando Oliveira, Gustavo Ramos de Rocha, Thales Lima Souza, Djair dos Santos de Silva, Maria Cristina Mattar da Franco, Octavio Luiz Grossi de Sa, Maria Fatima |
dc.subject.por.fl_str_mv |
Acanthoscelides obtectus Cysteine proteinase Propeptide Plant defense |
topic |
Acanthoscelides obtectus Cysteine proteinase Propeptide Plant defense |
dc.description.abstract.por.fl_txt_mv |
Acanthoscelides obtectus is a devastating storage insect pest capable of causing severe bean crop losses. In order to maintain their own development, insect pest larvae feed continuously, synthesizing efficient digestive enzymes. Among them, cysteine proteinases (CPs) are commonly produced as inactive precursors (procysteines), requiring a cleavage of the peptide proregion to become active. The proregion fits tightly into the active site of procysteines, efficiently preventing their activity. In this report, a CP cDNA (cpao) was isolated from A. obtectus midgut larvae. In silico studies indicated that the complete CP sequence contains a hydrophobic signal peptide, a prodomain and a conserved catalytic region. Moreover, the encoding cDNA contains 963 bp translating into a 321 residue protein, CPAo, which was expressed in E. coli, fused with thioredoxin. Enzymatic assays using the recombinant protein revealed that the enzyme was catalytically active, being able to cleave the synthetic substrate Z-Phe-Arg-7-AMC. Additionally, this report also focuses the cpao propeptide (PCPAo) subcloning and expression. The expressed propeptide efficiently inhibited CPAo, as well as digestive CP of other bean bruchids. Little or no activity was found against proteolytic enzymes of two other coleopterans: Rhyzopertha dominica and Anthonomus grandis. The data reported here indicate the possibility of endogenous propeptides as a novel strategy on bruchids control, which could be applicable to bean improvement programs. |
dc.description.version.pt_BR.fl_txt_mv |
Sim |
dc.description.status.pt_BR.fl_txt_mv |
Publicado |
description |
Acanthoscelides obtectus is a devastating storage insect pest capable of causing severe bean crop losses. In order to maintain their own development, insect pest larvae feed continuously, synthesizing efficient digestive enzymes. Among them, cysteine proteinases (CPs) are commonly produced as inactive precursors (procysteines), requiring a cleavage of the peptide proregion to become active. The proregion fits tightly into the active site of procysteines, efficiently preventing their activity. In this report, a CP cDNA (cpao) was isolated from A. obtectus midgut larvae. In silico studies indicated that the complete CP sequence contains a hydrophobic signal peptide, a prodomain and a conserved catalytic region. Moreover, the encoding cDNA contains 963 bp translating into a 321 residue protein, CPAo, which was expressed in E. coli, fused with thioredoxin. Enzymatic assays using the recombinant protein revealed that the enzyme was catalytically active, being able to cleave the synthetic substrate Z-Phe-Arg-7-AMC. Additionally, this report also focuses the cpao propeptide (PCPAo) subcloning and expression. The expressed propeptide efficiently inhibited CPAo, as well as digestive CP of other bean bruchids. Little or no activity was found against proteolytic enzymes of two other coleopterans: Rhyzopertha dominica and Anthonomus grandis. The data reported here indicate the possibility of endogenous propeptides as a novel strategy on bruchids control, which could be applicable to bean improvement programs. |
publishDate |
2007 |
dc.date.issued.fl_str_mv |
2007 |
dc.date.accessioned.fl_str_mv |
2016-10-10T03:52:40Z |
dc.date.available.fl_str_mv |
2016-10-10T03:52:40Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
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publishedVersion |
format |
article |
dc.identifier.citation.fl_str_mv |
SILVA, F. B. et al. Proregion of Acanthoscelides obtectus cysteine proteinase: a novel peptide with enhanced selectivity toward endogenous enzymes. Peptidies, v. 28, n. 6, p. 1292-1298, 2007. |
dc.identifier.uri.fl_str_mv |
http://twingo.ucb.br:8080/jspui/handle/10869/559 https://repositorio.ucb.br:9443/jspui/handle/123456789/7790 |
identifier_str_mv |
SILVA, F. B. et al. Proregion of Acanthoscelides obtectus cysteine proteinase: a novel peptide with enhanced selectivity toward endogenous enzymes. Peptidies, v. 28, n. 6, p. 1292-1298, 2007. |
url |
http://twingo.ucb.br:8080/jspui/handle/10869/559 https://repositorio.ucb.br:9443/jspui/handle/123456789/7790 |
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eng |
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eng |
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