Recombinant expression and characterization of a Xylella fastidiosa cysteine protease differentially expressed in a nonpathogenic strain

Detalhes bibliográficos
Autor(a) principal: Nogaroto, Viviane
Data de Publicação: 2006
Outros Autores: Tagliavini, Sandra A., Gianotti, Andreia, Mikawa, Angela, Barros, Nilana M. T., Puzer, Luciano, Carmona, Adriana K., Costa, Paulo I., Henrique-Silva, Flavio
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNESP
Texto Completo: http://dx.doi.org/10.1111/j.1574-6968.2006.00348.x
http://hdl.handle.net/11449/37403
Resumo: Xylella fastidiosa is a xylem-limited, Gram-negative bacterium responsible for citrus variegated chlorosis (CVC) in sweet oranges. In the present study, we present the recombinant expression, purification and characterization of an X. fastidiosa cysteine protease (dubbed Xylellain). The recombinant Xylellain ((HIS)Xylellain) was able to hydrolyze carbobenzoxy-Phe-Arg-7-amido-4-methylcoumarin (Z-FR-MCA) and carbobenzoxy-Arg-Arg-7-amido-4-methylcoumarin (Z-RR-MCA) with similar catalytic efficiencies, suggesting that this enzyme presents substrate specificity requirements similar to cathepsin B. The immunization of mice with (HIS)Xylellain provided us with antibodies, which recognized a protein of c. 31 kDa in the X. fastidiosa pathogenic strains 9a5c, and X. fastidiosa isolated from coffee plants. However, these antibodies recognized no protein in the nonpathogenic X. fastidiosa J1a12, suggesting the absence or low expression of this protein in the strain. These findings enabled us to identify Xylellain as a putative target for combating CVC and other diseases caused by X. fastidiosa strains.
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spelling Recombinant expression and characterization of a Xylella fastidiosa cysteine protease differentially expressed in a nonpathogenic strainXylella fastidiosacitrus variegated chlorosisphytopathogenrecombinant expressioncysteine proteasexylellainXylella fastidiosa is a xylem-limited, Gram-negative bacterium responsible for citrus variegated chlorosis (CVC) in sweet oranges. In the present study, we present the recombinant expression, purification and characterization of an X. fastidiosa cysteine protease (dubbed Xylellain). The recombinant Xylellain ((HIS)Xylellain) was able to hydrolyze carbobenzoxy-Phe-Arg-7-amido-4-methylcoumarin (Z-FR-MCA) and carbobenzoxy-Arg-Arg-7-amido-4-methylcoumarin (Z-RR-MCA) with similar catalytic efficiencies, suggesting that this enzyme presents substrate specificity requirements similar to cathepsin B. The immunization of mice with (HIS)Xylellain provided us with antibodies, which recognized a protein of c. 31 kDa in the X. fastidiosa pathogenic strains 9a5c, and X. fastidiosa isolated from coffee plants. However, these antibodies recognized no protein in the nonpathogenic X. fastidiosa J1a12, suggesting the absence or low expression of this protein in the strain. These findings enabled us to identify Xylellain as a putative target for combating CVC and other diseases caused by X. fastidiosa strains.Univ Fed Sao Carlos, Mol Biol Lab, DGE, BR-13565905 Sao Carlos, SP, BrazilUnifesp, Dept Biophys, São Paulo, SP, BrazilUNESP, Inst Chem, Araraquara, SP, BrazilSão Paulo State Univ, Fac Pharmaceut Sci, Dept Clin & Toxicol Anal, São Paulo, SP, BrazilUNESP, Inst Chem, Araraquara, SP, BrazilSão Paulo State Univ, Fac Pharmaceut Sci, Dept Clin & Toxicol Anal, São Paulo, SP, BrazilBlackwell PublishingUniversidade Federal de São Carlos (UFSCar)Universidade Federal de São Paulo (UNIFESP)Universidade Estadual Paulista (Unesp)Nogaroto, VivianeTagliavini, Sandra A.Gianotti, AndreiaMikawa, AngelaBarros, Nilana M. T.Puzer, LucianoCarmona, Adriana K.Costa, Paulo I.Henrique-Silva, Flavio2014-05-20T15:27:25Z2014-05-20T15:27:25Z2006-08-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article187-193application/pdfhttp://dx.doi.org/10.1111/j.1574-6968.2006.00348.xFems Microbiology Letters. Oxford: Blackwell Publishing, v. 261, n. 2, p. 187-193, 2006.0378-1097http://hdl.handle.net/11449/3740310.1111/j.1574-6968.2006.00348.xWOS:000239333800005WOS000239333800005.pdfWeb of Sciencereponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengFEMS Microbiology Letters1.7350,790info:eu-repo/semantics/openAccess2023-12-18T06:14:58Zoai:repositorio.unesp.br:11449/37403Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T20:37:31.585189Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false
dc.title.none.fl_str_mv Recombinant expression and characterization of a Xylella fastidiosa cysteine protease differentially expressed in a nonpathogenic strain
title Recombinant expression and characterization of a Xylella fastidiosa cysteine protease differentially expressed in a nonpathogenic strain
spellingShingle Recombinant expression and characterization of a Xylella fastidiosa cysteine protease differentially expressed in a nonpathogenic strain
Nogaroto, Viviane
Xylella fastidiosa
citrus variegated chlorosis
phytopathogen
recombinant expression
cysteine protease
xylellain
title_short Recombinant expression and characterization of a Xylella fastidiosa cysteine protease differentially expressed in a nonpathogenic strain
title_full Recombinant expression and characterization of a Xylella fastidiosa cysteine protease differentially expressed in a nonpathogenic strain
title_fullStr Recombinant expression and characterization of a Xylella fastidiosa cysteine protease differentially expressed in a nonpathogenic strain
title_full_unstemmed Recombinant expression and characterization of a Xylella fastidiosa cysteine protease differentially expressed in a nonpathogenic strain
title_sort Recombinant expression and characterization of a Xylella fastidiosa cysteine protease differentially expressed in a nonpathogenic strain
author Nogaroto, Viviane
author_facet Nogaroto, Viviane
Tagliavini, Sandra A.
Gianotti, Andreia
Mikawa, Angela
Barros, Nilana M. T.
Puzer, Luciano
Carmona, Adriana K.
Costa, Paulo I.
Henrique-Silva, Flavio
author_role author
author2 Tagliavini, Sandra A.
Gianotti, Andreia
Mikawa, Angela
Barros, Nilana M. T.
Puzer, Luciano
Carmona, Adriana K.
Costa, Paulo I.
Henrique-Silva, Flavio
author2_role author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade Federal de São Carlos (UFSCar)
Universidade Federal de São Paulo (UNIFESP)
Universidade Estadual Paulista (Unesp)
dc.contributor.author.fl_str_mv Nogaroto, Viviane
Tagliavini, Sandra A.
Gianotti, Andreia
Mikawa, Angela
Barros, Nilana M. T.
Puzer, Luciano
Carmona, Adriana K.
Costa, Paulo I.
Henrique-Silva, Flavio
dc.subject.por.fl_str_mv Xylella fastidiosa
citrus variegated chlorosis
phytopathogen
recombinant expression
cysteine protease
xylellain
topic Xylella fastidiosa
citrus variegated chlorosis
phytopathogen
recombinant expression
cysteine protease
xylellain
description Xylella fastidiosa is a xylem-limited, Gram-negative bacterium responsible for citrus variegated chlorosis (CVC) in sweet oranges. In the present study, we present the recombinant expression, purification and characterization of an X. fastidiosa cysteine protease (dubbed Xylellain). The recombinant Xylellain ((HIS)Xylellain) was able to hydrolyze carbobenzoxy-Phe-Arg-7-amido-4-methylcoumarin (Z-FR-MCA) and carbobenzoxy-Arg-Arg-7-amido-4-methylcoumarin (Z-RR-MCA) with similar catalytic efficiencies, suggesting that this enzyme presents substrate specificity requirements similar to cathepsin B. The immunization of mice with (HIS)Xylellain provided us with antibodies, which recognized a protein of c. 31 kDa in the X. fastidiosa pathogenic strains 9a5c, and X. fastidiosa isolated from coffee plants. However, these antibodies recognized no protein in the nonpathogenic X. fastidiosa J1a12, suggesting the absence or low expression of this protein in the strain. These findings enabled us to identify Xylellain as a putative target for combating CVC and other diseases caused by X. fastidiosa strains.
publishDate 2006
dc.date.none.fl_str_mv 2006-08-01
2014-05-20T15:27:25Z
2014-05-20T15:27:25Z
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 http://dx.doi.org/10.1111/j.1574-6968.2006.00348.x
Fems Microbiology Letters. Oxford: Blackwell Publishing, v. 261, n. 2, p. 187-193, 2006.
0378-1097
http://hdl.handle.net/11449/37403
10.1111/j.1574-6968.2006.00348.x
WOS:000239333800005
WOS000239333800005.pdf
url http://dx.doi.org/10.1111/j.1574-6968.2006.00348.x
http://hdl.handle.net/11449/37403
identifier_str_mv Fems Microbiology Letters. Oxford: Blackwell Publishing, v. 261, n. 2, p. 187-193, 2006.
0378-1097
10.1111/j.1574-6968.2006.00348.x
WOS:000239333800005
WOS000239333800005.pdf
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv FEMS Microbiology Letters
1.735
0,790
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv 187-193
application/pdf
dc.publisher.none.fl_str_mv Blackwell Publishing
publisher.none.fl_str_mv Blackwell Publishing
dc.source.none.fl_str_mv Web of Science
reponame:Repositório Institucional da UNESP
instname:Universidade Estadual Paulista (UNESP)
instacron:UNESP
instname_str Universidade Estadual Paulista (UNESP)
instacron_str UNESP
institution UNESP
reponame_str Repositório Institucional da UNESP
collection Repositório Institucional da UNESP
repository.name.fl_str_mv Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)
repository.mail.fl_str_mv
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