Proregion of Acanthoscelides obtectus cysteine proteinase: a novel peptide with enhanced selectivity toward endogenous enzymes

Bibliographic Details
Main Author: Silva, Francine Barbosa
Publication Date: 2007
Other Authors: 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
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.
id UCB-2_ecefe9fd56f1d9892bf6355245da075b
oai_identifier_str oai:200.214.135.189:123456789/7790
network_acronym_str UCB-2
network_name_str Repositório Institucional da UCB
spelling 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). No. of bitstreams: 5 Proregion of acanthoscelides obtectus cysteine proteinase.pdf: 1169326 bytes, checksum: eb5d3a1f34463cca4eb818a9ba1795ee (MD5) license_url: 52 bytes, checksum: 3d480ae6c91e310daba2020f8787d6f9 (MD5) license_text: 23851 bytes, checksum: 294cb7010cc40c47642971e073de3dba (MD5) license_rdf: 23892 bytes, checksum: afd5dad10b1d1e6dc10c8c5d25222c7a (MD5) license.txt: 1887 bytes, checksum: 445d1980f282ec865917de35a4c622f6 (MD5) Previous issue date: 2007SimPublicadoTextoRestrito UCBinfo:eu-repo/semantics/openAccessAcanthoscelides obtectusCysteine proteinasePropeptidePlant defenseProregion of Acanthoscelides obtectus cysteine proteinase: a novel peptide with enhanced selectivity toward endogenous enzymesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articlePeptidieshttp://www.sciencedirect.com/science?_ob=MiamiImageURL&_cid=271044&_user=1925346&_pii=S019697810700109X&_check=y&_origin=&_coverDate=30-Jun-2007&view=c&wchp=dGLzVlt-zSkWA&md5=a7918a440e6306c4e57a62b66c2fb13a/1-s2.0-S019697810700109X-main.pdfengreponame:Repositório Institucional da UCBinstname:Universidade Católica de Brasília (UCB)instacron:UCBORIGINALProregion of acanthoscelides obtectus cysteine proteinase.pdfapplication/pdf1169326https://200.214.135.178:9443/jspui/bitstream/123456789/7790/1/Proregion%20of%20acanthoscelides%20obtectus%20cysteine%20proteinase.pdfeb5d3a1f34463cca4eb818a9ba1795eeMD51CC-LICENSElicense_urlapplication/octet-stream52https://200.214.135.178:9443/jspui/bitstream/123456789/7790/2/license_url3d480ae6c91e310daba2020f8787d6f9MD52license_textapplication/octet-stream23851https://200.214.135.178:9443/jspui/bitstream/123456789/7790/3/license_text294cb7010cc40c47642971e073de3dbaMD53license_rdfapplication/octet-stream23892https://200.214.135.178:9443/jspui/bitstream/123456789/7790/4/license_rdfafd5dad10b1d1e6dc10c8c5d25222c7aMD54LICENSElicense.txttext/plain1887https://200.214.135.178:9443/jspui/bitstream/123456789/7790/5/license.txt445d1980f282ec865917de35a4c622f6MD55TEXTProregion of acanthoscelides obtectus cysteine proteinase.pdf.txtProregion of acanthoscelides obtectus cysteine proteinase.pdf.txtExtracted texttext/plain30442https://200.214.135.178:9443/jspui/bitstream/123456789/7790/6/Proregion%20of%20acanthoscelides%20obtectus%20cysteine%20proteinase.pdf.txt8805b098eac42cf8e69abb3bd318e285MD56123456789/77902017-01-17 15:10:55.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ório de Publicaçõeshttps://repositorio.ucb.br:9443/jspui/
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
status_str 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
dc.language.iso.fl_str_mv eng
language eng
dc.relation.publisherversion.pt_BR.fl_str_mv http://www.sciencedirect.com/science?_ob=MiamiImageURL&_cid=271044&_user=1925346&_pii=S019697810700109X&_check=y&_origin=&_coverDate=30-Jun-2007&view=c&wchp=dGLzVlt-zSkWA&md5=a7918a440e6306c4e57a62b66c2fb13a/1-s2.0-S019697810700109X-main.pdf
dc.rights.driver.fl_str_mv Restrito UCB
info:eu-repo/semantics/openAccess
rights_invalid_str_mv Restrito UCB
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv Texto
dc.source.none.fl_str_mv reponame:Repositório Institucional da UCB
instname:Universidade Católica de Brasília (UCB)
instacron:UCB
instname_str Universidade Católica de Brasília (UCB)
instacron_str UCB
institution UCB
reponame_str Repositório Institucional da UCB
collection Repositório Institucional da UCB
bitstream.url.fl_str_mv https://200.214.135.178:9443/jspui/bitstream/123456789/7790/1/Proregion%20of%20acanthoscelides%20obtectus%20cysteine%20proteinase.pdf
https://200.214.135.178:9443/jspui/bitstream/123456789/7790/2/license_url
https://200.214.135.178:9443/jspui/bitstream/123456789/7790/3/license_text
https://200.214.135.178:9443/jspui/bitstream/123456789/7790/4/license_rdf
https://200.214.135.178:9443/jspui/bitstream/123456789/7790/5/license.txt
https://200.214.135.178:9443/jspui/bitstream/123456789/7790/6/Proregion%20of%20acanthoscelides%20obtectus%20cysteine%20proteinase.pdf.txt
bitstream.checksum.fl_str_mv eb5d3a1f34463cca4eb818a9ba1795ee
3d480ae6c91e310daba2020f8787d6f9
294cb7010cc40c47642971e073de3dba
afd5dad10b1d1e6dc10c8c5d25222c7a
445d1980f282ec865917de35a4c622f6
8805b098eac42cf8e69abb3bd318e285
bitstream.checksumAlgorithm.fl_str_mv MD5
MD5
MD5
MD5
MD5
MD5
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1724829831261061120