Characterization of nitrophospholipid-peptide covalent adducts by electrospray tandem mass spectrometry: a first screening analysis using different instrumental platforms

Detalhes bibliográficos
Autor(a) principal: Montero-Bullon, Javier-Fernando
Data de Publicação: 2018
Outros Autores: Melo, Tânia, Domingues, Maria Rosário, Domingues, Pedro
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)
Texto Completo: http://hdl.handle.net/10773/27122
Resumo: Lipids are well‐known targets of reactive nitrogen species and this reaction leads to the formation of nitrated lipids that have been associated with anti‐inflammatory and cytoprotective effects. Nitro‐fatty acids (NO2‐FA) are highly electrophilic compounds that can form covalent adducts with proteins, leading to the formation of lipoxidation adducts, which modulate the protein structure and function. Nitrated phospholipids (NO2‐PL) have been detected recently in biological samples, but their biological effects are unknown, although similarly to what has been described for nitrated lipids, it has been hypothesized that they may react with peptides and proteins. In this study, in vitro biomimetic assays are used to synthetize adducts of nitrated POPC (NO2POPC), already detected in biological samples, and GSH peptide. The formation of NO2POPC‐GSH adducts is studied by ESI‐MS and MS2, using both low and high energy CID in different MS platforms: a LXQ linear ion trap, a Q‐TOF 2, and a Q‐Exactive Hybrid Quadrupole‐Orbitrap. Typical product ions observed under MS2 conditions are modified b, y, and C ions bearing NO2POPC covalently linked that unequivocally confirms the presence of the lipid‐peptide adduct. Typical loss of HNO2 is only observed in the MS2 of the mono‐charged precursor ions, [M+H]+. Product ions at m/z 184 or neutral loss of 183 Da are assigned as typical fragmentations that confirm the presence of the phosphatidylcholine. In summary, the characterization of nitro PL‐peptide adducts by MS and MS2 allows the identification of the structure and specific MS2 reporter ions to be used to pinpoint these adductions in biological systems.
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spelling Characterization of nitrophospholipid-peptide covalent adducts by electrospray tandem mass spectrometry: a first screening analysis using different instrumental platformsNitrationNitrophospholipidProtein adductsLipoxidationPost-translational modificationsLipids are well‐known targets of reactive nitrogen species and this reaction leads to the formation of nitrated lipids that have been associated with anti‐inflammatory and cytoprotective effects. Nitro‐fatty acids (NO2‐FA) are highly electrophilic compounds that can form covalent adducts with proteins, leading to the formation of lipoxidation adducts, which modulate the protein structure and function. Nitrated phospholipids (NO2‐PL) have been detected recently in biological samples, but their biological effects are unknown, although similarly to what has been described for nitrated lipids, it has been hypothesized that they may react with peptides and proteins. In this study, in vitro biomimetic assays are used to synthetize adducts of nitrated POPC (NO2POPC), already detected in biological samples, and GSH peptide. The formation of NO2POPC‐GSH adducts is studied by ESI‐MS and MS2, using both low and high energy CID in different MS platforms: a LXQ linear ion trap, a Q‐TOF 2, and a Q‐Exactive Hybrid Quadrupole‐Orbitrap. Typical product ions observed under MS2 conditions are modified b, y, and C ions bearing NO2POPC covalently linked that unequivocally confirms the presence of the lipid‐peptide adduct. Typical loss of HNO2 is only observed in the MS2 of the mono‐charged precursor ions, [M+H]+. Product ions at m/z 184 or neutral loss of 183 Da are assigned as typical fragmentations that confirm the presence of the phosphatidylcholine. In summary, the characterization of nitro PL‐peptide adducts by MS and MS2 allows the identification of the structure and specific MS2 reporter ions to be used to pinpoint these adductions in biological systems.Wiley2019-12-09T17:09:39Z2018-12-01T00:00:00Z2018-12info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10773/27122eng1438-769710.1002/ejlt.201800101Montero-Bullon, Javier-FernandoMelo, TâniaDomingues, Maria RosárioDomingues, Pedroinfo:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2024-02-22T11:52:33Zoai:ria.ua.pt:10773/27122Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-20T02:59:59.155074Repositório 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 Characterization of nitrophospholipid-peptide covalent adducts by electrospray tandem mass spectrometry: a first screening analysis using different instrumental platforms
title Characterization of nitrophospholipid-peptide covalent adducts by electrospray tandem mass spectrometry: a first screening analysis using different instrumental platforms
spellingShingle Characterization of nitrophospholipid-peptide covalent adducts by electrospray tandem mass spectrometry: a first screening analysis using different instrumental platforms
Montero-Bullon, Javier-Fernando
Nitration
Nitrophospholipid
Protein adducts
Lipoxidation
Post-translational modifications
title_short Characterization of nitrophospholipid-peptide covalent adducts by electrospray tandem mass spectrometry: a first screening analysis using different instrumental platforms
title_full Characterization of nitrophospholipid-peptide covalent adducts by electrospray tandem mass spectrometry: a first screening analysis using different instrumental platforms
title_fullStr Characterization of nitrophospholipid-peptide covalent adducts by electrospray tandem mass spectrometry: a first screening analysis using different instrumental platforms
title_full_unstemmed Characterization of nitrophospholipid-peptide covalent adducts by electrospray tandem mass spectrometry: a first screening analysis using different instrumental platforms
title_sort Characterization of nitrophospholipid-peptide covalent adducts by electrospray tandem mass spectrometry: a first screening analysis using different instrumental platforms
author Montero-Bullon, Javier-Fernando
author_facet Montero-Bullon, Javier-Fernando
Melo, Tânia
Domingues, Maria Rosário
Domingues, Pedro
author_role author
author2 Melo, Tânia
Domingues, Maria Rosário
Domingues, Pedro
author2_role author
author
author
dc.contributor.author.fl_str_mv Montero-Bullon, Javier-Fernando
Melo, Tânia
Domingues, Maria Rosário
Domingues, Pedro
dc.subject.por.fl_str_mv Nitration
Nitrophospholipid
Protein adducts
Lipoxidation
Post-translational modifications
topic Nitration
Nitrophospholipid
Protein adducts
Lipoxidation
Post-translational modifications
description Lipids are well‐known targets of reactive nitrogen species and this reaction leads to the formation of nitrated lipids that have been associated with anti‐inflammatory and cytoprotective effects. Nitro‐fatty acids (NO2‐FA) are highly electrophilic compounds that can form covalent adducts with proteins, leading to the formation of lipoxidation adducts, which modulate the protein structure and function. Nitrated phospholipids (NO2‐PL) have been detected recently in biological samples, but their biological effects are unknown, although similarly to what has been described for nitrated lipids, it has been hypothesized that they may react with peptides and proteins. In this study, in vitro biomimetic assays are used to synthetize adducts of nitrated POPC (NO2POPC), already detected in biological samples, and GSH peptide. The formation of NO2POPC‐GSH adducts is studied by ESI‐MS and MS2, using both low and high energy CID in different MS platforms: a LXQ linear ion trap, a Q‐TOF 2, and a Q‐Exactive Hybrid Quadrupole‐Orbitrap. Typical product ions observed under MS2 conditions are modified b, y, and C ions bearing NO2POPC covalently linked that unequivocally confirms the presence of the lipid‐peptide adduct. Typical loss of HNO2 is only observed in the MS2 of the mono‐charged precursor ions, [M+H]+. Product ions at m/z 184 or neutral loss of 183 Da are assigned as typical fragmentations that confirm the presence of the phosphatidylcholine. In summary, the characterization of nitro PL‐peptide adducts by MS and MS2 allows the identification of the structure and specific MS2 reporter ions to be used to pinpoint these adductions in biological systems.
publishDate 2018
dc.date.none.fl_str_mv 2018-12-01T00:00:00Z
2018-12
2019-12-09T17:09:39Z
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
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status_str publishedVersion
dc.identifier.uri.fl_str_mv http://hdl.handle.net/10773/27122
url http://hdl.handle.net/10773/27122
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 1438-7697
10.1002/ejlt.201800101
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dc.publisher.none.fl_str_mv Wiley
publisher.none.fl_str_mv Wiley
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