The two main endoproteases present in dark-induced senescent wheat leaves are distinct subtilisin-like proteases

Autores
Roberts, Irma; Di Bernardo, Maria Susana; Barneix, Atilio José
Año de publicación
2006
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
We have previously reported the occurrence of two serine endoproteases (referred as P1 and P2) in dark-induced senescent wheat (Triticum aestivum L.) leaves. P1 enzyme was already purified and identified as a subtilisin-like serine endoprotease. In this paper, we demonstrate by Western blot analysis of extracts obtained from dark-induced senescent leaves that an antiserum raised against P1 was able to recognise a second protein band of 78 kDa which corresponded to P2 activity. This result suggested that both enzymes must be structurally related. Therefore, we purified and characterized P2 activity. Its biochemical and physical properties (inhibition by chymostatin and PMSF, broad pH range of activity, thermostability and ability to hydrolyse Suc-AAPF-pNA) allowed its classification as chymotripsin-like protease. Samples of purified P1 and P2 were submitted to MALDI-TOF mass spectrometry analysis. Comparison of the peptide maps obtained clearly showed that P1 and P2 do not share peptides in common, although P2 was also identified as a subtilisin-like serine protease. Western blot analysis demonstrated that P1 was induced in non-detached dark-induced senescent leaves but was not detectable in senescent leaves promoted by nitrogen (N) deprivation. In contrast, P2 was already present in non-senescent leaves and its expression increased by darkness as well as by N starvation. These results indicate that, despite their biochemical and structural similarities, both enzymes are probably involved in different physiological roles.
Fil: Roberts, Irma. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales; Argentina
Fil: Di Bernardo, Maria Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales; Argentina
Fil: Barneix, Atilio José. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales; Argentina
Materia
SUBTILISIN
SERIN PROTEASE
SENESCENCE
WHEAT
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/244124
Acceder
id CONICETDig_627cd4c804170319e5bee01e7fd79e67
oai_identifier_str oai:ri.conicet.gov.ar:11336/244124
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling The two main endoproteases present in dark-induced senescent wheat leaves are distinct subtilisin-like proteasesRoberts, IrmaDi Bernardo, Maria SusanaBarneix, Atilio JoséSUBTILISINSERIN PROTEASESENESCENCEWHEAThttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1We have previously reported the occurrence of two serine endoproteases (referred as P1 and P2) in dark-induced senescent wheat (Triticum aestivum L.) leaves. P1 enzyme was already purified and identified as a subtilisin-like serine endoprotease. In this paper, we demonstrate by Western blot analysis of extracts obtained from dark-induced senescent leaves that an antiserum raised against P1 was able to recognise a second protein band of 78 kDa which corresponded to P2 activity. This result suggested that both enzymes must be structurally related. Therefore, we purified and characterized P2 activity. Its biochemical and physical properties (inhibition by chymostatin and PMSF, broad pH range of activity, thermostability and ability to hydrolyse Suc-AAPF-pNA) allowed its classification as chymotripsin-like protease. Samples of purified P1 and P2 were submitted to MALDI-TOF mass spectrometry analysis. Comparison of the peptide maps obtained clearly showed that P1 and P2 do not share peptides in common, although P2 was also identified as a subtilisin-like serine protease. Western blot analysis demonstrated that P1 was induced in non-detached dark-induced senescent leaves but was not detectable in senescent leaves promoted by nitrogen (N) deprivation. In contrast, P2 was already present in non-senescent leaves and its expression increased by darkness as well as by N starvation. These results indicate that, despite their biochemical and structural similarities, both enzymes are probably involved in different physiological roles.Fil: Roberts, Irma. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales; ArgentinaFil: Di Bernardo, Maria Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales; ArgentinaFil: Barneix, Atilio José. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales; ArgentinaSpringer2006-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/244124Roberts, Irma; Di Bernardo, Maria Susana; Barneix, Atilio José; The two main endoproteases present in dark-induced senescent wheat leaves are distinct subtilisin-like proteases; Springer; Planta; 224; 6; 6-2006; 1437-14470032-0935CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://link.springer.com/article/10.1007/s00425-006-0312-2info:eu-repo/semantics/altIdentifier/doi/10.1007/s00425-006-0312-2info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:06:43Zoai:ri.conicet.gov.ar:11336/244124instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 10:06:43.338CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv The two main endoproteases present in dark-induced senescent wheat leaves are distinct subtilisin-like proteases
title The two main endoproteases present in dark-induced senescent wheat leaves are distinct subtilisin-like proteases
spellingShingle The two main endoproteases present in dark-induced senescent wheat leaves are distinct subtilisin-like proteases
Roberts, Irma
SUBTILISIN
SERIN PROTEASE
SENESCENCE
WHEAT
title_short The two main endoproteases present in dark-induced senescent wheat leaves are distinct subtilisin-like proteases
title_full The two main endoproteases present in dark-induced senescent wheat leaves are distinct subtilisin-like proteases
title_fullStr The two main endoproteases present in dark-induced senescent wheat leaves are distinct subtilisin-like proteases
title_full_unstemmed The two main endoproteases present in dark-induced senescent wheat leaves are distinct subtilisin-like proteases
title_sort The two main endoproteases present in dark-induced senescent wheat leaves are distinct subtilisin-like proteases
dc.creator.none.fl_str_mv Roberts, Irma
Di Bernardo, Maria Susana
Barneix, Atilio José
author Roberts, Irma
author_facet Roberts, Irma
Di Bernardo, Maria Susana
Barneix, Atilio José
author_role author
author2 Di Bernardo, Maria Susana
Barneix, Atilio José
author2_role author
author
dc.subject.none.fl_str_mv SUBTILISIN
SERIN PROTEASE
SENESCENCE
WHEAT
topic SUBTILISIN
SERIN PROTEASE
SENESCENCE
WHEAT
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv We have previously reported the occurrence of two serine endoproteases (referred as P1 and P2) in dark-induced senescent wheat (Triticum aestivum L.) leaves. P1 enzyme was already purified and identified as a subtilisin-like serine endoprotease. In this paper, we demonstrate by Western blot analysis of extracts obtained from dark-induced senescent leaves that an antiserum raised against P1 was able to recognise a second protein band of 78 kDa which corresponded to P2 activity. This result suggested that both enzymes must be structurally related. Therefore, we purified and characterized P2 activity. Its biochemical and physical properties (inhibition by chymostatin and PMSF, broad pH range of activity, thermostability and ability to hydrolyse Suc-AAPF-pNA) allowed its classification as chymotripsin-like protease. Samples of purified P1 and P2 were submitted to MALDI-TOF mass spectrometry analysis. Comparison of the peptide maps obtained clearly showed that P1 and P2 do not share peptides in common, although P2 was also identified as a subtilisin-like serine protease. Western blot analysis demonstrated that P1 was induced in non-detached dark-induced senescent leaves but was not detectable in senescent leaves promoted by nitrogen (N) deprivation. In contrast, P2 was already present in non-senescent leaves and its expression increased by darkness as well as by N starvation. These results indicate that, despite their biochemical and structural similarities, both enzymes are probably involved in different physiological roles.
Fil: Roberts, Irma. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales; Argentina
Fil: Di Bernardo, Maria Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales; Argentina
Fil: Barneix, Atilio José. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales; Argentina
description We have previously reported the occurrence of two serine endoproteases (referred as P1 and P2) in dark-induced senescent wheat (Triticum aestivum L.) leaves. P1 enzyme was already purified and identified as a subtilisin-like serine endoprotease. In this paper, we demonstrate by Western blot analysis of extracts obtained from dark-induced senescent leaves that an antiserum raised against P1 was able to recognise a second protein band of 78 kDa which corresponded to P2 activity. This result suggested that both enzymes must be structurally related. Therefore, we purified and characterized P2 activity. Its biochemical and physical properties (inhibition by chymostatin and PMSF, broad pH range of activity, thermostability and ability to hydrolyse Suc-AAPF-pNA) allowed its classification as chymotripsin-like protease. Samples of purified P1 and P2 were submitted to MALDI-TOF mass spectrometry analysis. Comparison of the peptide maps obtained clearly showed that P1 and P2 do not share peptides in common, although P2 was also identified as a subtilisin-like serine protease. Western blot analysis demonstrated that P1 was induced in non-detached dark-induced senescent leaves but was not detectable in senescent leaves promoted by nitrogen (N) deprivation. In contrast, P2 was already present in non-senescent leaves and its expression increased by darkness as well as by N starvation. These results indicate that, despite their biochemical and structural similarities, both enzymes are probably involved in different physiological roles.
publishDate 2006
dc.date.none.fl_str_mv 2006-06
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/244124
Roberts, Irma; Di Bernardo, Maria Susana; Barneix, Atilio José; The two main endoproteases present in dark-induced senescent wheat leaves are distinct subtilisin-like proteases; Springer; Planta; 224; 6; 6-2006; 1437-1447
0032-0935
CONICET Digital
CONICET
url http://hdl.handle.net/11336/244124
identifier_str_mv Roberts, Irma; Di Bernardo, Maria Susana; Barneix, Atilio José; The two main endoproteases present in dark-induced senescent wheat leaves are distinct subtilisin-like proteases; Springer; Planta; 224; 6; 6-2006; 1437-1447
0032-0935
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://link.springer.com/article/10.1007/s00425-006-0312-2
info:eu-repo/semantics/altIdentifier/doi/10.1007/s00425-006-0312-2
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Springer
publisher.none.fl_str_mv Springer
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.13397

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