Recombinant expression and characterization of a Xylella fastidiosa cysteine protease differentially expressed in a nonpathogenic strain
Autor(a) principal: | |
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Data de Publicação: | 2006 |
Outros Autores: | , , , , , , , |
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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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 |
|
_version_ |
1808129229410271232 |