Characterization of human sperm protamine proteoforms through a combination of top-down and bottom-up mass spectrometry approaches

Protamine 1 (P1) and protamine 2 (P2) family are extremely basic, sperm-specific proteins, packing 85-95% of the paternal DNA. P1 is synthesized as a mature form, whereas P2 components (HP2, HP3, and HP4) arise from the proteolysis of the precursor (pre-P2). Due to the particular protamine physical-...

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Detalhes bibliográficos
Autores: Soler Ventura, Ada, Gay i Marín, Marina, Jodar Bifet, Meritxell, Vilanova, Mar, Castillo Corullón, Judit, Arauz-Garofalo, Gianluca, Villarreal, Laura, Ballescà, Josep Lluís, Vilaseca, Marta, Oliva Virgili, Rafael
Formato: artículo
Estado:Versión aceptada para publicación
Fecha de publicación:2020
País:España
Recursos:Universidad de Barcelona
Repositorio:Dipòsit Digital de la UB
OAI Identifier:oai:diposit.ub.edu:2445/218071
Acesso em linha:https://hdl.handle.net/2445/218071
Access Level:acceso abierto
Palavra-chave:Espermatozoides
Fosforilació
Esterilitat masculina
Protamines
Proteòmica
Spermatozoa
Phosphorylation
Male sterility
Proteomics
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spelling Characterization of human sperm protamine proteoforms through a combination of top-down and bottom-up mass spectrometry approachesSoler Ventura, AdaGay i Marín, MarinaJodar Bifet, MeritxellVilanova, MarCastillo Corullón, JuditArauz-Garofalo, GianlucaVillarreal, LauraBallescà, Josep LluísVilaseca, MartaOliva Virgili, RafaelEspermatozoidesFosforilacióEsterilitat masculinaProtaminesProteòmicaSpermatozoaPhosphorylationMale sterilityProtaminesProteomicsProtamine 1 (P1) and protamine 2 (P2) family are extremely basic, sperm-specific proteins, packing 85-95% of the paternal DNA. P1 is synthesized as a mature form, whereas P2 components (HP2, HP3, and HP4) arise from the proteolysis of the precursor (pre-P2). Due to the particular protamine physical-chemical properties, their identification by standardized bottom-up mass spectrometry (MS) strategies is not straightforward. Therefore, the aim of this study was to identify the sperm protamine proteoforms profile, including their post-translational modifications, in normozoospermic individuals using two complementary strategies, a top-down MS approach and a proteinase-K-digestion-based bottom-up MS approach. By top-down MS, described and novel truncated P1 and pre-P2 proteoforms were identified. Intact P1, pre-P2, and P2 mature proteoforms and their phosphorylation pattern were also detected. Additionally, a +61 Da modification in different proteoforms was observed. By the bottom-up MS approach, phosphorylated residues for pre-P2, as well as the new P2 isoform 2, which is not annotated in the UniProtKB database, were revealed. Implementing these strategies in comparative studies of different infertile phenotypes, together with the evaluation of P1/P2 and pre-P2/P2 MS-derived ratios, would permit determining specific alterations in the protamine proteoforms and elucidate the role of phosphorylation/dephosphorylation dynamics in male fertility.American Chemical Society2020info:eu-repo/semantics/articleinfo:eu-repo/semantics/acceptedVersionapplication/pdfhttps://hdl.handle.net/2445/218071Articles publicats en revistes (Biomedicina)reponame:Dipòsit Digital de la UBinstname:Universidad de BarcelonaInglésVersió postprint del document publicat a: https://doi.org/10.1021/acs.jproteome.9b00499Journal of Proteome Research, 2020, vol. 19, num.1, p. 221-237https://doi.org/10.1021/acs.jproteome.9b00499(c) American Chemical Society, 2020info:eu-repo/semantics/openAccessoai:diposit.ub.edu:2445/2180712026-05-27T06:46:51Z
dc.title.none.fl_str_mv Characterization of human sperm protamine proteoforms through a combination of top-down and bottom-up mass spectrometry approaches
title Characterization of human sperm protamine proteoforms through a combination of top-down and bottom-up mass spectrometry approaches
spellingShingle Characterization of human sperm protamine proteoforms through a combination of top-down and bottom-up mass spectrometry approaches
Soler Ventura, Ada
Espermatozoides
Fosforilació
Esterilitat masculina
Protamines
Proteòmica
Spermatozoa
Phosphorylation
Male sterility
Protamines
Proteomics
title_short Characterization of human sperm protamine proteoforms through a combination of top-down and bottom-up mass spectrometry approaches
title_full Characterization of human sperm protamine proteoforms through a combination of top-down and bottom-up mass spectrometry approaches
title_fullStr Characterization of human sperm protamine proteoforms through a combination of top-down and bottom-up mass spectrometry approaches
title_full_unstemmed Characterization of human sperm protamine proteoforms through a combination of top-down and bottom-up mass spectrometry approaches
title_sort Characterization of human sperm protamine proteoforms through a combination of top-down and bottom-up mass spectrometry approaches
dc.creator.none.fl_str_mv Soler Ventura, Ada
Gay i Marín, Marina
Jodar Bifet, Meritxell
Vilanova, Mar
Castillo Corullón, Judit
Arauz-Garofalo, Gianluca
Villarreal, Laura
Ballescà, Josep Lluís
Vilaseca, Marta
Oliva Virgili, Rafael
author Soler Ventura, Ada
author_facet Soler Ventura, Ada
Gay i Marín, Marina
Jodar Bifet, Meritxell
Vilanova, Mar
Castillo Corullón, Judit
Arauz-Garofalo, Gianluca
Villarreal, Laura
Ballescà, Josep Lluís
Vilaseca, Marta
Oliva Virgili, Rafael
author_role author
author2 Gay i Marín, Marina
Jodar Bifet, Meritxell
Vilanova, Mar
Castillo Corullón, Judit
Arauz-Garofalo, Gianluca
Villarreal, Laura
Ballescà, Josep Lluís
Vilaseca, Marta
Oliva Virgili, Rafael
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Espermatozoides
Fosforilació
Esterilitat masculina
Protamines
Proteòmica
Spermatozoa
Phosphorylation
Male sterility
Protamines
Proteomics
topic Espermatozoides
Fosforilació
Esterilitat masculina
Protamines
Proteòmica
Spermatozoa
Phosphorylation
Male sterility
Protamines
Proteomics
description Protamine 1 (P1) and protamine 2 (P2) family are extremely basic, sperm-specific proteins, packing 85-95% of the paternal DNA. P1 is synthesized as a mature form, whereas P2 components (HP2, HP3, and HP4) arise from the proteolysis of the precursor (pre-P2). Due to the particular protamine physical-chemical properties, their identification by standardized bottom-up mass spectrometry (MS) strategies is not straightforward. Therefore, the aim of this study was to identify the sperm protamine proteoforms profile, including their post-translational modifications, in normozoospermic individuals using two complementary strategies, a top-down MS approach and a proteinase-K-digestion-based bottom-up MS approach. By top-down MS, described and novel truncated P1 and pre-P2 proteoforms were identified. Intact P1, pre-P2, and P2 mature proteoforms and their phosphorylation pattern were also detected. Additionally, a +61 Da modification in different proteoforms was observed. By the bottom-up MS approach, phosphorylated residues for pre-P2, as well as the new P2 isoform 2, which is not annotated in the UniProtKB database, were revealed. Implementing these strategies in comparative studies of different infertile phenotypes, together with the evaluation of P1/P2 and pre-P2/P2 MS-derived ratios, would permit determining specific alterations in the protamine proteoforms and elucidate the role of phosphorylation/dephosphorylation dynamics in male fertility.
publishDate 2020
dc.date.none.fl_str_mv 2020
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/acceptedVersion
format article
status_str acceptedVersion
dc.identifier.none.fl_str_mv https://hdl.handle.net/2445/218071
url https://hdl.handle.net/2445/218071
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.relation.none.fl_str_mv Versió postprint del document publicat a: https://doi.org/10.1021/acs.jproteome.9b00499
Journal of Proteome Research, 2020, vol. 19, num.1, p. 221-237
https://doi.org/10.1021/acs.jproteome.9b00499
dc.rights.none.fl_str_mv (c) American Chemical Society, 2020
info:eu-repo/semantics/openAccess
rights_invalid_str_mv (c) American Chemical Society, 2020
eu_rights_str_mv openAccess
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv American Chemical Society
publisher.none.fl_str_mv American Chemical Society
dc.source.none.fl_str_mv Articles publicats en revistes (Biomedicina)
reponame:Dipòsit Digital de la UB
instname:Universidad de Barcelona
instname_str Universidad de Barcelona
reponame_str Dipòsit Digital de la UB
collection Dipòsit Digital de la UB
repository.name.fl_str_mv
repository.mail.fl_str_mv
_version_ 1869406397795401728
score 15,811543

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