PRSS38 Is a Novel Sperm Serine Protease Involved in Human and Mouse Fertilization

Autores
Manjon, Ania Antonella; Verón, Gustavo Luis; Vitale, Rosario; Stegmayer, Georgina; Gonzalez Echeverria Raffo, Fernanda; Lane, Lydie; Vazquez, Monica Hebe
Año de publicación
2025
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Sperm proteases are involved in several gamete interaction events leading to fertilization.This report presents a detailed analysis on the expression and localization of serine proteasePRSS38 in human and in mouse spermatozoa and its involvement in fertilization-related events, using bioinformatics, cellular, biochemical, molecular and functionalapproaches. Bioinformatics analyses included genomics and data analysis, prediction ofprotein subcellular localization and post-translational modifications, Self-OrganizingMaps (SOMs) unsupervised training with other serine proteases, protein modeling (AlphaFold) and genetic variant analysis. For cellular, biochemical and functional studies,human semen samples were obtained from healthy normozoospermic volunteers, andcauda epididymal sperm were collected from adult Balb-c/C57 mice. PRSS38 presencewas detected in human and mouse sperm protein extracts by Western immunobloting.Sperm PRSS38 subcellular localization was determined by fluorescence immunocytochemistry.Human sperm-oocyte interaction events were assessed by means of themouse Cumulus Penetration Assay (CPA) using mouse COCs, the human HemizonaAssay (HZA), and the ZP-free hamster egg Sperm Penetration Assay (SPA). Mousesperm-oocyte interactions were evaluated by means of in vitro fertilization (IVF) withCOCs and denuded oocytes. PRSS38 is proposed to be a GPI-anchored serine protease(active site: His-100, Asp-150 and Ser-245) based on bioinformatics analyses. Usingcommercial antibodies, protein forms of the expected Mr (human: 31 kDa; mouse: 32 and24 kDa) were specifically immunodetected in protein sperm extracts. Immunocytochemicalanalysis revealed a specific PRSS38 signal in the human sperm acrosomal region,equatorial segment, and flagellum. Mouse sperm PRSS38 was immunolocalized inthe equatorial segment and hook. Human sperm preincubation with specific antibodiesresulted in inhibition (P<0.05) of CPA, HZA, and SPA. Mouse sperm preincubation withPRSS38 antibodies impaired (P<0.05) homologous IVF using COCs and denuded oocytes.Genetic variants affecting residues involved in the GPI anchor and the catalytic triadwere found in individuals from the general population whose PRSS38 protease functioncould be altered. This study provides, for the first time, an integrated analysis of PRSS38in human and mouse sperm, contributing our understanding of mammalian fertilization and male infertility.
Fil: Manjon, Ania Antonella. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Verón, Gustavo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Vitale, Rosario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Investigación en Señales, Sistemas e Inteligencia Computacional. Universidad Nacional del Litoral. Facultad de Ingeniería y Ciencias Hídricas. Instituto de Investigación en Señales, Sistemas e Inteligencia Computacional; Argentina
Fil: Stegmayer, Georgina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Investigación en Señales, Sistemas e Inteligencia Computacional. Universidad Nacional del Litoral. Facultad de Ingeniería y Ciencias Hídricas. Instituto de Investigación en Señales, Sistemas e Inteligencia Computacional; Argentina
Fil: Gonzalez Echeverria Raffo, Fernanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Lane, Lydie. No especifíca;
Fil: Vazquez, Monica Hebe. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Materia
human reproduction
fertility
self organizing maps
clustering
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/278361
Acceder
id CONICETDig_3f865a5df91c9cfc20e04bbde52588b1
oai_identifier_str oai:ri.conicet.gov.ar:11336/278361
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling PRSS38 Is a Novel Sperm Serine Protease Involved in Human and Mouse FertilizationManjon, Ania AntonellaVerón, Gustavo LuisVitale, RosarioStegmayer, GeorginaGonzalez Echeverria Raffo, FernandaLane, LydieVazquez, Monica Hebehuman reproductionfertilityself organizing mapsclusteringhttps://purl.org/becyt/ford/1.2https://purl.org/becyt/ford/1Sperm proteases are involved in several gamete interaction events leading to fertilization.This report presents a detailed analysis on the expression and localization of serine proteasePRSS38 in human and in mouse spermatozoa and its involvement in fertilization-related events, using bioinformatics, cellular, biochemical, molecular and functionalapproaches. Bioinformatics analyses included genomics and data analysis, prediction ofprotein subcellular localization and post-translational modifications, Self-OrganizingMaps (SOMs) unsupervised training with other serine proteases, protein modeling (AlphaFold) and genetic variant analysis. For cellular, biochemical and functional studies,human semen samples were obtained from healthy normozoospermic volunteers, andcauda epididymal sperm were collected from adult Balb-c/C57 mice. PRSS38 presencewas detected in human and mouse sperm protein extracts by Western immunobloting.Sperm PRSS38 subcellular localization was determined by fluorescence immunocytochemistry.Human sperm-oocyte interaction events were assessed by means of themouse Cumulus Penetration Assay (CPA) using mouse COCs, the human HemizonaAssay (HZA), and the ZP-free hamster egg Sperm Penetration Assay (SPA). Mousesperm-oocyte interactions were evaluated by means of in vitro fertilization (IVF) withCOCs and denuded oocytes. PRSS38 is proposed to be a GPI-anchored serine protease(active site: His-100, Asp-150 and Ser-245) based on bioinformatics analyses. Usingcommercial antibodies, protein forms of the expected Mr (human: 31 kDa; mouse: 32 and24 kDa) were specifically immunodetected in protein sperm extracts. Immunocytochemicalanalysis revealed a specific PRSS38 signal in the human sperm acrosomal region,equatorial segment, and flagellum. Mouse sperm PRSS38 was immunolocalized inthe equatorial segment and hook. Human sperm preincubation with specific antibodiesresulted in inhibition (P<0.05) of CPA, HZA, and SPA. Mouse sperm preincubation withPRSS38 antibodies impaired (P<0.05) homologous IVF using COCs and denuded oocytes.Genetic variants affecting residues involved in the GPI anchor and the catalytic triadwere found in individuals from the general population whose PRSS38 protease functioncould be altered. This study provides, for the first time, an integrated analysis of PRSS38in human and mouse sperm, contributing our understanding of mammalian fertilization and male infertility.Fil: Manjon, Ania Antonella. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Verón, Gustavo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Vitale, Rosario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Investigación en Señales, Sistemas e Inteligencia Computacional. Universidad Nacional del Litoral. Facultad de Ingeniería y Ciencias Hídricas. Instituto de Investigación en Señales, Sistemas e Inteligencia Computacional; ArgentinaFil: Stegmayer, Georgina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Investigación en Señales, Sistemas e Inteligencia Computacional. Universidad Nacional del Litoral. Facultad de Ingeniería y Ciencias Hídricas. Instituto de Investigación en Señales, Sistemas e Inteligencia Computacional; ArgentinaFil: Gonzalez Echeverria Raffo, Fernanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaFil: Lane, Lydie. No especifíca;Fil: Vazquez, Monica Hebe. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; ArgentinaMolecular Diversity Preservation International2025-10info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/278361Manjon, Ania Antonella; Verón, Gustavo Luis; Vitale, Rosario; Stegmayer, Georgina; Gonzalez Echeverria Raffo, Fernanda; et al.; PRSS38 Is a Novel Sperm Serine Protease Involved in Human and Mouse Fertilization; Molecular Diversity Preservation International; International Journal of Molecular Sciences; 26; 23; 10-2025; 1-251422-0067CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.mdpi.com/1422-0067/26/23/11680info:eu-repo/semantics/altIdentifier/doi/10.3390/ijms262311680info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2026-01-08T12:47:51Zoai:ri.conicet.gov.ar:11336/278361instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982026-01-08 12:47:51.678CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv PRSS38 Is a Novel Sperm Serine Protease Involved in Human and Mouse Fertilization
title PRSS38 Is a Novel Sperm Serine Protease Involved in Human and Mouse Fertilization
spellingShingle PRSS38 Is a Novel Sperm Serine Protease Involved in Human and Mouse Fertilization
Manjon, Ania Antonella
human reproduction
fertility
self organizing maps
clustering
title_short PRSS38 Is a Novel Sperm Serine Protease Involved in Human and Mouse Fertilization
title_full PRSS38 Is a Novel Sperm Serine Protease Involved in Human and Mouse Fertilization
title_fullStr PRSS38 Is a Novel Sperm Serine Protease Involved in Human and Mouse Fertilization
title_full_unstemmed PRSS38 Is a Novel Sperm Serine Protease Involved in Human and Mouse Fertilization
title_sort PRSS38 Is a Novel Sperm Serine Protease Involved in Human and Mouse Fertilization
dc.creator.none.fl_str_mv Manjon, Ania Antonella
Verón, Gustavo Luis
Vitale, Rosario
Stegmayer, Georgina
Gonzalez Echeverria Raffo, Fernanda
Lane, Lydie
Vazquez, Monica Hebe
author Manjon, Ania Antonella
author_facet Manjon, Ania Antonella
Verón, Gustavo Luis
Vitale, Rosario
Stegmayer, Georgina
Gonzalez Echeverria Raffo, Fernanda
Lane, Lydie
Vazquez, Monica Hebe
author_role author
author2 Verón, Gustavo Luis
Vitale, Rosario
Stegmayer, Georgina
Gonzalez Echeverria Raffo, Fernanda
Lane, Lydie
Vazquez, Monica Hebe
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv human reproduction
fertility
self organizing maps
clustering
topic human reproduction
fertility
self organizing maps
clustering
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.2
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Sperm proteases are involved in several gamete interaction events leading to fertilization.This report presents a detailed analysis on the expression and localization of serine proteasePRSS38 in human and in mouse spermatozoa and its involvement in fertilization-related events, using bioinformatics, cellular, biochemical, molecular and functionalapproaches. Bioinformatics analyses included genomics and data analysis, prediction ofprotein subcellular localization and post-translational modifications, Self-OrganizingMaps (SOMs) unsupervised training with other serine proteases, protein modeling (AlphaFold) and genetic variant analysis. For cellular, biochemical and functional studies,human semen samples were obtained from healthy normozoospermic volunteers, andcauda epididymal sperm were collected from adult Balb-c/C57 mice. PRSS38 presencewas detected in human and mouse sperm protein extracts by Western immunobloting.Sperm PRSS38 subcellular localization was determined by fluorescence immunocytochemistry.Human sperm-oocyte interaction events were assessed by means of themouse Cumulus Penetration Assay (CPA) using mouse COCs, the human HemizonaAssay (HZA), and the ZP-free hamster egg Sperm Penetration Assay (SPA). Mousesperm-oocyte interactions were evaluated by means of in vitro fertilization (IVF) withCOCs and denuded oocytes. PRSS38 is proposed to be a GPI-anchored serine protease(active site: His-100, Asp-150 and Ser-245) based on bioinformatics analyses. Usingcommercial antibodies, protein forms of the expected Mr (human: 31 kDa; mouse: 32 and24 kDa) were specifically immunodetected in protein sperm extracts. Immunocytochemicalanalysis revealed a specific PRSS38 signal in the human sperm acrosomal region,equatorial segment, and flagellum. Mouse sperm PRSS38 was immunolocalized inthe equatorial segment and hook. Human sperm preincubation with specific antibodiesresulted in inhibition (P<0.05) of CPA, HZA, and SPA. Mouse sperm preincubation withPRSS38 antibodies impaired (P<0.05) homologous IVF using COCs and denuded oocytes.Genetic variants affecting residues involved in the GPI anchor and the catalytic triadwere found in individuals from the general population whose PRSS38 protease functioncould be altered. This study provides, for the first time, an integrated analysis of PRSS38in human and mouse sperm, contributing our understanding of mammalian fertilization and male infertility.
Fil: Manjon, Ania Antonella. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Verón, Gustavo Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Vitale, Rosario. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Investigación en Señales, Sistemas e Inteligencia Computacional. Universidad Nacional del Litoral. Facultad de Ingeniería y Ciencias Hídricas. Instituto de Investigación en Señales, Sistemas e Inteligencia Computacional; Argentina
Fil: Stegmayer, Georgina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Santa Fe. Instituto de Investigación en Señales, Sistemas e Inteligencia Computacional. Universidad Nacional del Litoral. Facultad de Ingeniería y Ciencias Hídricas. Instituto de Investigación en Señales, Sistemas e Inteligencia Computacional; Argentina
Fil: Gonzalez Echeverria Raffo, Fernanda. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
Fil: Lane, Lydie. No especifíca;
Fil: Vazquez, Monica Hebe. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Biología y Medicina Experimental. Fundación de Instituto de Biología y Medicina Experimental. Instituto de Biología y Medicina Experimental; Argentina
description Sperm proteases are involved in several gamete interaction events leading to fertilization.This report presents a detailed analysis on the expression and localization of serine proteasePRSS38 in human and in mouse spermatozoa and its involvement in fertilization-related events, using bioinformatics, cellular, biochemical, molecular and functionalapproaches. Bioinformatics analyses included genomics and data analysis, prediction ofprotein subcellular localization and post-translational modifications, Self-OrganizingMaps (SOMs) unsupervised training with other serine proteases, protein modeling (AlphaFold) and genetic variant analysis. For cellular, biochemical and functional studies,human semen samples were obtained from healthy normozoospermic volunteers, andcauda epididymal sperm were collected from adult Balb-c/C57 mice. PRSS38 presencewas detected in human and mouse sperm protein extracts by Western immunobloting.Sperm PRSS38 subcellular localization was determined by fluorescence immunocytochemistry.Human sperm-oocyte interaction events were assessed by means of themouse Cumulus Penetration Assay (CPA) using mouse COCs, the human HemizonaAssay (HZA), and the ZP-free hamster egg Sperm Penetration Assay (SPA). Mousesperm-oocyte interactions were evaluated by means of in vitro fertilization (IVF) withCOCs and denuded oocytes. PRSS38 is proposed to be a GPI-anchored serine protease(active site: His-100, Asp-150 and Ser-245) based on bioinformatics analyses. Usingcommercial antibodies, protein forms of the expected Mr (human: 31 kDa; mouse: 32 and24 kDa) were specifically immunodetected in protein sperm extracts. Immunocytochemicalanalysis revealed a specific PRSS38 signal in the human sperm acrosomal region,equatorial segment, and flagellum. Mouse sperm PRSS38 was immunolocalized inthe equatorial segment and hook. Human sperm preincubation with specific antibodiesresulted in inhibition (P<0.05) of CPA, HZA, and SPA. Mouse sperm preincubation withPRSS38 antibodies impaired (P<0.05) homologous IVF using COCs and denuded oocytes.Genetic variants affecting residues involved in the GPI anchor and the catalytic triadwere found in individuals from the general population whose PRSS38 protease functioncould be altered. This study provides, for the first time, an integrated analysis of PRSS38in human and mouse sperm, contributing our understanding of mammalian fertilization and male infertility.
publishDate 2025
dc.date.none.fl_str_mv 2025-10
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/278361
Manjon, Ania Antonella; Verón, Gustavo Luis; Vitale, Rosario; Stegmayer, Georgina; Gonzalez Echeverria Raffo, Fernanda; et al.; PRSS38 Is a Novel Sperm Serine Protease Involved in Human and Mouse Fertilization; Molecular Diversity Preservation International; International Journal of Molecular Sciences; 26; 23; 10-2025; 1-25
1422-0067
CONICET Digital
CONICET
url http://hdl.handle.net/11336/278361
identifier_str_mv Manjon, Ania Antonella; Verón, Gustavo Luis; Vitale, Rosario; Stegmayer, Georgina; Gonzalez Echeverria Raffo, Fernanda; et al.; PRSS38 Is a Novel Sperm Serine Protease Involved in Human and Mouse Fertilization; Molecular Diversity Preservation International; International Journal of Molecular Sciences; 26; 23; 10-2025; 1-25
1422-0067
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.mdpi.com/1422-0067/26/23/11680
info:eu-repo/semantics/altIdentifier/doi/10.3390/ijms262311680
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Molecular Diversity Preservation International
publisher.none.fl_str_mv Molecular Diversity Preservation International
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.25844

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