Improved protein identification efficiency by mass spectrometry using N-terminal chemical derivatization of peptides from Angiostrongylus costaricensis, a nematode with unknown genome

Detalhes bibliográficos
Autor(a) principal: León, Ileana R.
Data de Publicação: 2007
Outros Autores: Ferreira, Ana G. C. Neves, Valente, Richard H., Mota, Ester Maria, Lenzi, Henrique Leonel, Perales, Jonas
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da FIOCRUZ (ARCA)
Texto Completo: https://www.arca.fiocruz.br/handle/icict/5964
Resumo: Acknowledgements This study was supported by grants from Conselho Nacional de Desenvolvimento Científico and Tecnológico (CNPq), Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) and Fundação Oswaldo Cruz-PDTIS, Brazil. IRL thanks Instituto Oswaldo Cruz for her Master's scholarship.
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spelling León, Ileana R.Ferreira, Ana G. C. NevesValente, Richard H.Mota, Ester MariaLenzi, Henrique LeonelPerales, Jonas2012-12-12T15:21:02Z2012-12-12T15:21:02Z2007LEON, I. R. et al. Improved protein identification efficiency by mass spectrometry using N-terminal chemical derivatization of peptides from Angiostrongylus costaricensis, a nematode with unknown genome. Journal of Mass Spectrometry, v. 42, p. 781-792, 2007.1076-5174https://www.arca.fiocruz.br/handle/icict/596410.1002/jms.1214Acknowledgements This study was supported by grants from Conselho Nacional de Desenvolvimento Científico and Tecnológico (CNPq), Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) and Fundação Oswaldo Cruz-PDTIS, Brazil. IRL thanks Instituto Oswaldo Cruz for her Master's scholarship.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Fisiologia e Farmacodinâmica. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Fisiologia e Farmacodinâmica. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Fisiologia e Farmacodinâmica. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Patologia. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Patologia. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto Oswaldo Cruz. Departamento de Fisiologia e Farmacodinâmica. Rio de Janeiro, RJ, Brasil.Matrix-assisted laser desorption ionization (MALDI), Peptide Mass Fingerprinting (PMF) and MALDI-MS/MS ion search (using MASCOT) have become the preferred methods for high-throughput identification of proteins. Unfortunately, PMF can be ambiguous, mainly when the genome of the organism under investigation is unknown and the quality of spectra generated is poor and does not allow confident identification. The post-source decay (PSD) fragmentation of singly charged tryptic peptide ions generated by MALDI-TOF/TOF typically results in low fragmentation efficiency and/or complex spectra, including backbone fragmentation ions (series b and y), internal fragmentation etc. Interpreting these data either manually and/or using de novo sequencing software can frequently be a challenge. To overcome this limitation when studying the proteome of adult Angiostrongylus costaricensis, a nematode with unknown genome, we have used chemical N-terminal derivatization of the tryptic peptides with 4-sulfophenyl isothiocyanate (SPITC) prior to MALDI-TOF/TOF MS. This methodology has recently been reported to enhance the quality of MALDI-TOF/TOF-PSD data, allowing the obtainment of complete sequence of most of the peptides and thus facilitating de novo peptide sequencing. Our approach, consisting of SPITC derivatization along with manual spectra interpretation and Blast analysis, was able to positively identify 76% of analyzed samples, whereas MASCOT analysis of derivatized samples, MASCOT analysis of nonderivatized samples and PMF of nonderivatized samples yielded only 35, 41 and 12% positive identifications, respectively. Moreover, de novo sequencing of SPITC modified peptides resulted in protein sequences not available in NCBInr database paving the way to the discovery of new protein molecules.engWileyImproved protein identification efficiency by mass spectrometry using N-terminal chemical derivatization of peptides from Angiostrongylus costaricensis, a nematode with unknown genomeinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleSPITCMALDI TOF/TOFPSDde novo sequencingAngiostrongylus costaricensisSequência de AminoácidosAngiostrongylus, químicaProteínas de HelmintoDados de Sequência MolecularMapeamento de Peptídeosinfo:eu-repo/semantics/openAccessreponame:Repositório Institucional da FIOCRUZ (ARCA)instname:Fundação Oswaldo Cruz (FIOCRUZ)instacron:FIOCRUZLICENSElicense.txtlicense.txttext/plain; charset=utf-81949https://www.arca.fiocruz.br/bitstream/icict/5964/2/license.txtaad6e5ff39a8b3723a1981fb3bcd1becMD52TEXTImproved protein identification efficiency by mass.htm.txtImproved protein 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dc.title.pt_BR.fl_str_mv Improved protein identification efficiency by mass spectrometry using N-terminal chemical derivatization of peptides from Angiostrongylus costaricensis, a nematode with unknown genome
title Improved protein identification efficiency by mass spectrometry using N-terminal chemical derivatization of peptides from Angiostrongylus costaricensis, a nematode with unknown genome
spellingShingle Improved protein identification efficiency by mass spectrometry using N-terminal chemical derivatization of peptides from Angiostrongylus costaricensis, a nematode with unknown genome
León, Ileana R.
SPITC
MALDI TOF/TOF
PSD
de novo sequencing
Angiostrongylus costaricensis
Sequência de Aminoácidos
Angiostrongylus, química
Proteínas de Helminto
Dados de Sequência Molecular
Mapeamento de Peptídeos
title_short Improved protein identification efficiency by mass spectrometry using N-terminal chemical derivatization of peptides from Angiostrongylus costaricensis, a nematode with unknown genome
title_full Improved protein identification efficiency by mass spectrometry using N-terminal chemical derivatization of peptides from Angiostrongylus costaricensis, a nematode with unknown genome
title_fullStr Improved protein identification efficiency by mass spectrometry using N-terminal chemical derivatization of peptides from Angiostrongylus costaricensis, a nematode with unknown genome
title_full_unstemmed Improved protein identification efficiency by mass spectrometry using N-terminal chemical derivatization of peptides from Angiostrongylus costaricensis, a nematode with unknown genome
title_sort Improved protein identification efficiency by mass spectrometry using N-terminal chemical derivatization of peptides from Angiostrongylus costaricensis, a nematode with unknown genome
author León, Ileana R.
author_facet León, Ileana R.
Ferreira, Ana G. C. Neves
Valente, Richard H.
Mota, Ester Maria
Lenzi, Henrique Leonel
Perales, Jonas
author_role author
author2 Ferreira, Ana G. C. Neves
Valente, Richard H.
Mota, Ester Maria
Lenzi, Henrique Leonel
Perales, Jonas
author2_role author
author
author
author
author
dc.contributor.author.fl_str_mv León, Ileana R.
Ferreira, Ana G. C. Neves
Valente, Richard H.
Mota, Ester Maria
Lenzi, Henrique Leonel
Perales, Jonas
dc.subject.en.pt_BR.fl_str_mv SPITC
MALDI TOF/TOF
PSD
de novo sequencing
Angiostrongylus costaricensis
topic SPITC
MALDI TOF/TOF
PSD
de novo sequencing
Angiostrongylus costaricensis
Sequência de Aminoácidos
Angiostrongylus, química
Proteínas de Helminto
Dados de Sequência Molecular
Mapeamento de Peptídeos
dc.subject.decs.pt_BR.fl_str_mv Sequência de Aminoácidos
Angiostrongylus, química
Proteínas de Helminto
Dados de Sequência Molecular
Mapeamento de Peptídeos
description Acknowledgements This study was supported by grants from Conselho Nacional de Desenvolvimento Científico and Tecnológico (CNPq), Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (FAPERJ) and Fundação Oswaldo Cruz-PDTIS, Brazil. IRL thanks Instituto Oswaldo Cruz for her Master's scholarship.
publishDate 2007
dc.date.issued.fl_str_mv 2007
dc.date.accessioned.fl_str_mv 2012-12-12T15:21:02Z
dc.date.available.fl_str_mv 2012-12-12T15:21:02Z
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.citation.fl_str_mv LEON, I. R. et al. Improved protein identification efficiency by mass spectrometry using N-terminal chemical derivatization of peptides from Angiostrongylus costaricensis, a nematode with unknown genome. Journal of Mass Spectrometry, v. 42, p. 781-792, 2007.
dc.identifier.uri.fl_str_mv https://www.arca.fiocruz.br/handle/icict/5964
dc.identifier.issn.none.fl_str_mv 1076-5174
dc.identifier.doi.none.fl_str_mv 10.1002/jms.1214
identifier_str_mv LEON, I. R. et al. Improved protein identification efficiency by mass spectrometry using N-terminal chemical derivatization of peptides from Angiostrongylus costaricensis, a nematode with unknown genome. Journal of Mass Spectrometry, v. 42, p. 781-792, 2007.
1076-5174
10.1002/jms.1214
url https://www.arca.fiocruz.br/handle/icict/5964
dc.language.iso.fl_str_mv eng
language eng
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv Wiley
publisher.none.fl_str_mv Wiley
dc.source.none.fl_str_mv reponame:Repositório Institucional da FIOCRUZ (ARCA)
instname:Fundação Oswaldo Cruz (FIOCRUZ)
instacron:FIOCRUZ
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instacron_str FIOCRUZ
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