Structural and enzymatic characterization of Peruvianin‑I, the first germin-like protein with proteolytic activity

Detalhes bibliográficos
Autor(a) principal: Cruz, Wallace T. da
Data de Publicação: 2019
Outros Autores: Bezerra, Eduardo H. S., Grangeiro, Thalles B., Lopes, Jose L. S., Silva, Maria Z. R., Ramos, Márcio Viana, Rocha, Bruno A. M., Oliveira, Jefferson S., Freitas, Deborah C., Freitas, Cleverson D. T.
Tipo de documento: Artigo
Idioma: por
Título da fonte: Repositório Institucional da Universidade Federal do Ceará (UFC)
Texto Completo: http://www.repositorio.ufc.br/handle/riufc/63486
Resumo: The germin-like protein (GLP) purified from Thevetia peruviana, Peruvianin-I, is the only one described as having proteolytic activity. Therefore, the goal of this study was to investigate the structural features responsible for its enzymatic activity. Although the amino acid sequence of Peruvianin-I showed high identity with other GLPs, it exhibited punctual mutations, which were responsible for the absence of oxalate oxidase activity. The phylogenetic analysis showed that Peruvianin-I does not belong to any classification of GLP subfamilies. Moreover, Peruvianin-I contains a catalytic triad found in all plant cysteine peptidases. Molecular docking simulations confirmed the role of the catalytic triad in its proteolytic activity. Synchrotron radiation circular dichroism assays confirmed that Peruvianin-I was stable at pH ranging from 5.0 to 8.0 and that it presented significant structural changes only above 60 °C. The addition of iodoacetamide caused changes in its native conformation, but only a slight effect was observed after adding a reducing agent. This study reports an unusual protein with germin-like structure, lacking typical oxalate oxidase activity. Instead, the proteolytic activity observed suggests that the protein is a cysteine peptidase. These structural peculiarities make Peruvianin‑I an interesting model for further understanding of the action of laticifer fluids in plant defense.
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spelling Structural and enzymatic characterization of Peruvianin‑I, the first germin-like protein with proteolytic activityGLPLaticiferOxalate oxidaseThe germin-like protein (GLP) purified from Thevetia peruviana, Peruvianin-I, is the only one described as having proteolytic activity. Therefore, the goal of this study was to investigate the structural features responsible for its enzymatic activity. Although the amino acid sequence of Peruvianin-I showed high identity with other GLPs, it exhibited punctual mutations, which were responsible for the absence of oxalate oxidase activity. The phylogenetic analysis showed that Peruvianin-I does not belong to any classification of GLP subfamilies. Moreover, Peruvianin-I contains a catalytic triad found in all plant cysteine peptidases. Molecular docking simulations confirmed the role of the catalytic triad in its proteolytic activity. Synchrotron radiation circular dichroism assays confirmed that Peruvianin-I was stable at pH ranging from 5.0 to 8.0 and that it presented significant structural changes only above 60 °C. The addition of iodoacetamide caused changes in its native conformation, but only a slight effect was observed after adding a reducing agent. This study reports an unusual protein with germin-like structure, lacking typical oxalate oxidase activity. Instead, the proteolytic activity observed suggests that the protein is a cysteine peptidase. These structural peculiarities make Peruvianin‑I an interesting model for further understanding of the action of laticifer fluids in plant defense.International Journal of Biological Macromolecules2022-01-13T17:23:19Z2022-01-13T17:23:19Z2019info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfCRUZ, Wallace T. da et al. Structural and enzymatic characterization of Peruvianin‑I, the first germin-like protein with proteolytic activity. International Journal of Biological Macromolecules, [s. l.], v. 126, p. 1167-1176, 2019.http://www.repositorio.ufc.br/handle/riufc/63486Cruz, Wallace T. daBezerra, Eduardo H. S.Grangeiro, Thalles B.Lopes, Jose L. S.Silva, Maria Z. R.Ramos, Márcio VianaRocha, Bruno A. M.Oliveira, Jefferson S.Freitas, Deborah C.Freitas, Cleverson D. T.info:eu-repo/semantics/openAccessporreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFC2023-10-10T19:33:28Zoai:repositorio.ufc.br:riufc/63486Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2024-09-11T18:26:40.354945Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false
dc.title.none.fl_str_mv Structural and enzymatic characterization of Peruvianin‑I, the first germin-like protein with proteolytic activity
title Structural and enzymatic characterization of Peruvianin‑I, the first germin-like protein with proteolytic activity
spellingShingle Structural and enzymatic characterization of Peruvianin‑I, the first germin-like protein with proteolytic activity
Cruz, Wallace T. da
GLP
Laticifer
Oxalate oxidase
title_short Structural and enzymatic characterization of Peruvianin‑I, the first germin-like protein with proteolytic activity
title_full Structural and enzymatic characterization of Peruvianin‑I, the first germin-like protein with proteolytic activity
title_fullStr Structural and enzymatic characterization of Peruvianin‑I, the first germin-like protein with proteolytic activity
title_full_unstemmed Structural and enzymatic characterization of Peruvianin‑I, the first germin-like protein with proteolytic activity
title_sort Structural and enzymatic characterization of Peruvianin‑I, the first germin-like protein with proteolytic activity
author Cruz, Wallace T. da
author_facet Cruz, Wallace T. da
Bezerra, Eduardo H. S.
Grangeiro, Thalles B.
Lopes, Jose L. S.
Silva, Maria Z. R.
Ramos, Márcio Viana
Rocha, Bruno A. M.
Oliveira, Jefferson S.
Freitas, Deborah C.
Freitas, Cleverson D. T.
author_role author
author2 Bezerra, Eduardo H. S.
Grangeiro, Thalles B.
Lopes, Jose L. S.
Silva, Maria Z. R.
Ramos, Márcio Viana
Rocha, Bruno A. M.
Oliveira, Jefferson S.
Freitas, Deborah C.
Freitas, Cleverson D. T.
author2_role author
author
author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Cruz, Wallace T. da
Bezerra, Eduardo H. S.
Grangeiro, Thalles B.
Lopes, Jose L. S.
Silva, Maria Z. R.
Ramos, Márcio Viana
Rocha, Bruno A. M.
Oliveira, Jefferson S.
Freitas, Deborah C.
Freitas, Cleverson D. T.
dc.subject.por.fl_str_mv GLP
Laticifer
Oxalate oxidase
topic GLP
Laticifer
Oxalate oxidase
description The germin-like protein (GLP) purified from Thevetia peruviana, Peruvianin-I, is the only one described as having proteolytic activity. Therefore, the goal of this study was to investigate the structural features responsible for its enzymatic activity. Although the amino acid sequence of Peruvianin-I showed high identity with other GLPs, it exhibited punctual mutations, which were responsible for the absence of oxalate oxidase activity. The phylogenetic analysis showed that Peruvianin-I does not belong to any classification of GLP subfamilies. Moreover, Peruvianin-I contains a catalytic triad found in all plant cysteine peptidases. Molecular docking simulations confirmed the role of the catalytic triad in its proteolytic activity. Synchrotron radiation circular dichroism assays confirmed that Peruvianin-I was stable at pH ranging from 5.0 to 8.0 and that it presented significant structural changes only above 60 °C. The addition of iodoacetamide caused changes in its native conformation, but only a slight effect was observed after adding a reducing agent. This study reports an unusual protein with germin-like structure, lacking typical oxalate oxidase activity. Instead, the proteolytic activity observed suggests that the protein is a cysteine peptidase. These structural peculiarities make Peruvianin‑I an interesting model for further understanding of the action of laticifer fluids in plant defense.
publishDate 2019
dc.date.none.fl_str_mv 2019
2022-01-13T17:23:19Z
2022-01-13T17:23:19Z
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.uri.fl_str_mv CRUZ, Wallace T. da et al. Structural and enzymatic characterization of Peruvianin‑I, the first germin-like protein with proteolytic activity. International Journal of Biological Macromolecules, [s. l.], v. 126, p. 1167-1176, 2019.
http://www.repositorio.ufc.br/handle/riufc/63486
identifier_str_mv CRUZ, Wallace T. da et al. Structural and enzymatic characterization of Peruvianin‑I, the first germin-like protein with proteolytic activity. International Journal of Biological Macromolecules, [s. l.], v. 126, p. 1167-1176, 2019.
url http://www.repositorio.ufc.br/handle/riufc/63486
dc.language.iso.fl_str_mv por
language por
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv International Journal of Biological Macromolecules
publisher.none.fl_str_mv International Journal of Biological Macromolecules
dc.source.none.fl_str_mv reponame:Repositório Institucional da Universidade Federal do Ceará (UFC)
instname:Universidade Federal do Ceará (UFC)
instacron:UFC
instname_str Universidade Federal do Ceará (UFC)
instacron_str UFC
institution UFC
reponame_str Repositório Institucional da Universidade Federal do Ceará (UFC)
collection Repositório Institucional da Universidade Federal do Ceará (UFC)
repository.name.fl_str_mv Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)
repository.mail.fl_str_mv bu@ufc.br || repositorio@ufc.br
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