Mitochondria proteome profiling: a comparative analysis between gel- and gel-free approaches
Autor(a) principal: | |
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Data de Publicação: | 2013 |
Outros Autores: | , , , , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10400.18/2221 |
Resumo: | Mitochondrial proteomics emerged aiming to disclose the dynamics of mitochondria under various pathophysiological conditions. In the present study we investigated the relative merits of gel-based (2DE and SDS-LC) and gel-free (2D-LC) protein separation approaches and protein identification algorithms (Mascot and Paragon) in the proteome profiling of mitochondria isolated from cultured fibroblasts, a sample traditionally used for diagnosis purposes. Combining data retrieved from 2DE, 2D-LC and SDS-LC and search methods, a total of 696 non-redundant proteins were identified. An overlap of only 19% between the proteins identified by the three different methods was observed when Mascot and Paragon were used. Regarding protein ID, a consistency in the number of identified proteins per sample was noticed for 2DE approach. Independent of the methodological approach chosen, it was noticed that the predominance in mitochondria of hydrophilic proteins with 20-50 kDa and pI 5-6 and 8-9; however, 2D-LC and SDS-LC allowed the enrichment of proteins with a mass below 30 kDa and of basic proteins with pI values above 8. In conclusion, data from the present study highlight the power of integrating different separation technologies and protein identification algorithms. |
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Mitochondria proteome profiling: a comparative analysis between gel- and gel-free approachesDoenças GenéticasProteoma MitocondrialElectrophoresisGel-basedGel-freeLC-MS/MSPTMsProteomicsMitochondrial proteomics emerged aiming to disclose the dynamics of mitochondria under various pathophysiological conditions. In the present study we investigated the relative merits of gel-based (2DE and SDS-LC) and gel-free (2D-LC) protein separation approaches and protein identification algorithms (Mascot and Paragon) in the proteome profiling of mitochondria isolated from cultured fibroblasts, a sample traditionally used for diagnosis purposes. Combining data retrieved from 2DE, 2D-LC and SDS-LC and search methods, a total of 696 non-redundant proteins were identified. An overlap of only 19% between the proteins identified by the three different methods was observed when Mascot and Paragon were used. Regarding protein ID, a consistency in the number of identified proteins per sample was noticed for 2DE approach. Independent of the methodological approach chosen, it was noticed that the predominance in mitochondria of hydrophilic proteins with 20-50 kDa and pI 5-6 and 8-9; however, 2D-LC and SDS-LC allowed the enrichment of proteins with a mass below 30 kDa and of basic proteins with pI values above 8. In conclusion, data from the present study highlight the power of integrating different separation technologies and protein identification algorithms.ElsevierRepositório Científico do Instituto Nacional de SaúdeFerreira, R.Rocha, H.Almeida, V.Padrão, A.L.Santa, C.Vilarinho, L.Amado, F.Vitorino, R.2014-04-03T12:14:20Z2013-10-152013-10-15T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10400.18/2221engTalanta. 2013 Oct 15;115:277-83. doi: 10.1016/j.talanta.2013.04.026. Epub 2013 Apr 220039-9140doi: 10.1016/j.talanta.2013.04.026info:eu-repo/semantics/embargoedAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-20T15:39:10Zoai:repositorio.insa.pt:10400.18/2221Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T18:37:15.994038Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Mitochondria proteome profiling: a comparative analysis between gel- and gel-free approaches |
title |
Mitochondria proteome profiling: a comparative analysis between gel- and gel-free approaches |
spellingShingle |
Mitochondria proteome profiling: a comparative analysis between gel- and gel-free approaches Ferreira, R. Doenças Genéticas Proteoma Mitocondrial Electrophoresis Gel-based Gel-free LC-MS/MS PTMs Proteomics |
title_short |
Mitochondria proteome profiling: a comparative analysis between gel- and gel-free approaches |
title_full |
Mitochondria proteome profiling: a comparative analysis between gel- and gel-free approaches |
title_fullStr |
Mitochondria proteome profiling: a comparative analysis between gel- and gel-free approaches |
title_full_unstemmed |
Mitochondria proteome profiling: a comparative analysis between gel- and gel-free approaches |
title_sort |
Mitochondria proteome profiling: a comparative analysis between gel- and gel-free approaches |
author |
Ferreira, R. |
author_facet |
Ferreira, R. Rocha, H. Almeida, V. Padrão, A.L. Santa, C. Vilarinho, L. Amado, F. Vitorino, R. |
author_role |
author |
author2 |
Rocha, H. Almeida, V. Padrão, A.L. Santa, C. Vilarinho, L. Amado, F. Vitorino, R. |
author2_role |
author author author author author author author |
dc.contributor.none.fl_str_mv |
Repositório Científico do Instituto Nacional de Saúde |
dc.contributor.author.fl_str_mv |
Ferreira, R. Rocha, H. Almeida, V. Padrão, A.L. Santa, C. Vilarinho, L. Amado, F. Vitorino, R. |
dc.subject.por.fl_str_mv |
Doenças Genéticas Proteoma Mitocondrial Electrophoresis Gel-based Gel-free LC-MS/MS PTMs Proteomics |
topic |
Doenças Genéticas Proteoma Mitocondrial Electrophoresis Gel-based Gel-free LC-MS/MS PTMs Proteomics |
description |
Mitochondrial proteomics emerged aiming to disclose the dynamics of mitochondria under various pathophysiological conditions. In the present study we investigated the relative merits of gel-based (2DE and SDS-LC) and gel-free (2D-LC) protein separation approaches and protein identification algorithms (Mascot and Paragon) in the proteome profiling of mitochondria isolated from cultured fibroblasts, a sample traditionally used for diagnosis purposes. Combining data retrieved from 2DE, 2D-LC and SDS-LC and search methods, a total of 696 non-redundant proteins were identified. An overlap of only 19% between the proteins identified by the three different methods was observed when Mascot and Paragon were used. Regarding protein ID, a consistency in the number of identified proteins per sample was noticed for 2DE approach. Independent of the methodological approach chosen, it was noticed that the predominance in mitochondria of hydrophilic proteins with 20-50 kDa and pI 5-6 and 8-9; however, 2D-LC and SDS-LC allowed the enrichment of proteins with a mass below 30 kDa and of basic proteins with pI values above 8. In conclusion, data from the present study highlight the power of integrating different separation technologies and protein identification algorithms. |
publishDate |
2013 |
dc.date.none.fl_str_mv |
2013-10-15 2013-10-15T00:00:00Z 2014-04-03T12:14:20Z |
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://hdl.handle.net/10400.18/2221 |
url |
http://hdl.handle.net/10400.18/2221 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Talanta. 2013 Oct 15;115:277-83. doi: 10.1016/j.talanta.2013.04.026. Epub 2013 Apr 22 0039-9140 doi: 10.1016/j.talanta.2013.04.026 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/embargoedAccess |
eu_rights_str_mv |
embargoedAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Elsevier |
publisher.none.fl_str_mv |
Elsevier |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
institution |
RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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1799132106644783104 |