Study of the antibacterial peptide P8.1: Effect on anionic vesicles using spectroscopic techniques

Autores
Jimenez, Luis Emanuel; Alvarez, Rosa Maria Susana; Maffia, Paulo Cesar; Hollmann, Axel
Año de publicación
2025
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Antimicrobial peptides (AMPs) are promising complements to antibiotics, yet their membrane-level actions remain incompletely understood. In this work, we characterized how the “de novo” cationic AMP P8.1 interacts with anionic lipid bilayers composed of DPPG (gel phase) or DLPG (fluid phase) using zeta potential, tryptophan and Laurdan fluorescence, Raman microscopy, and a carboxyfluorescein (CF) leakage assay. P8.1 bound both lipids electrostatically, reduced zeta potential, and increased large unilamellar vesicles (LUVs) size. Binding kinetics were faster on DPPG multilamellar vesicles, whereas Trp fluorescence assays showed deeper insertion in DLPG (larger Trp blue-shift and lower acrylamide quenching). Laurdan generalized polarization (GP) increased in DLPG but not in DPPG, indicating reduced water access and higher local order in fluid bilayers. Raman spectra revealed diminished phosphate-band intensity in both systems and, in DLPG, a decreased gauche/trans ratio and narrower 1300 cm-1 band consistent with tighter acyl-chain packing. Difference spectra further showed an amide I shift of P8.1, supporting a random-coil to α-helix transition upon binding to lipids. Finally, P8.1 induced ∼80% CF leakage in DLPG LUVs within minutes. Together, the data indicate that P8.1 engages phosphate groups of lipids and then modulates bilayer structure in a phase- and mechanics-dependent manner—rigidifying short-chain, fluid DLPG and minimally perturbing gel-phase DPPG—providing mechanistic insight relevant to antibacterial activity.
Fil: Jimenez, Luis Emanuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto de Química del Noroeste. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Química del Noroeste; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Química Física; Argentina. Universidad Nacional de Tucumán. Facultad de Ciencias Naturales e Instituto Miguel Lillo; Argentina
Fil: Alvarez, Rosa Maria Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto de Química del Noroeste. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Química del Noroeste; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Química Física; Argentina
Fil: Maffia, Paulo Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Laboratorio de Biotecnologia y Microbiologia Aplicada ; Secretaria de Investigacion ; Universidad Nacional de Hurlingham;
Fil: Hollmann, Axel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Centro de Investigación en Biofísica Aplicada y Alimentos. - Universidad Nacional de Santiago del Estero. Centro de Investigación en Biofísica Aplicada y Alimentos; Argentina
Materia
Antimicrobial Peptide
Lipid Phase State Membrane
DLPG
DPPG
Nivel de accesibilidad
acceso embargado
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:11336/277472
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/277472
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Study of the antibacterial peptide P8.1: Effect on anionic vesicles using spectroscopic techniquesJimenez, Luis EmanuelAlvarez, Rosa Maria SusanaMaffia, Paulo CesarHollmann, AxelAntimicrobial PeptideLipid Phase State MembraneDLPGDPPGhttps://purl.org/becyt/ford/1.4https://purl.org/becyt/ford/1Antimicrobial peptides (AMPs) are promising complements to antibiotics, yet their membrane-level actions remain incompletely understood. In this work, we characterized how the “de novo” cationic AMP P8.1 interacts with anionic lipid bilayers composed of DPPG (gel phase) or DLPG (fluid phase) using zeta potential, tryptophan and Laurdan fluorescence, Raman microscopy, and a carboxyfluorescein (CF) leakage assay. P8.1 bound both lipids electrostatically, reduced zeta potential, and increased large unilamellar vesicles (LUVs) size. Binding kinetics were faster on DPPG multilamellar vesicles, whereas Trp fluorescence assays showed deeper insertion in DLPG (larger Trp blue-shift and lower acrylamide quenching). Laurdan generalized polarization (GP) increased in DLPG but not in DPPG, indicating reduced water access and higher local order in fluid bilayers. Raman spectra revealed diminished phosphate-band intensity in both systems and, in DLPG, a decreased gauche/trans ratio and narrower 1300 cm-1 band consistent with tighter acyl-chain packing. Difference spectra further showed an amide I shift of P8.1, supporting a random-coil to α-helix transition upon binding to lipids. Finally, P8.1 induced ∼80% CF leakage in DLPG LUVs within minutes. Together, the data indicate that P8.1 engages phosphate groups of lipids and then modulates bilayer structure in a phase- and mechanics-dependent manner—rigidifying short-chain, fluid DLPG and minimally perturbing gel-phase DPPG—providing mechanistic insight relevant to antibacterial activity.Fil: Jimenez, Luis Emanuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto de Química del Noroeste. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Química del Noroeste; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Química Física; Argentina. Universidad Nacional de Tucumán. Facultad de Ciencias Naturales e Instituto Miguel Lillo; ArgentinaFil: Alvarez, Rosa Maria Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto de Química del Noroeste. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Química del Noroeste; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Química Física; ArgentinaFil: Maffia, Paulo Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Laboratorio de Biotecnologia y Microbiologia Aplicada ; Secretaria de Investigacion ; Universidad Nacional de Hurlingham;Fil: Hollmann, Axel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Centro de Investigación en Biofísica Aplicada y Alimentos. - Universidad Nacional de Santiago del Estero. Centro de Investigación en Biofísica Aplicada y Alimentos; ArgentinaElsevier Science Inc.2025-12info:eu-repo/date/embargoEnd/2026-06-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/277472Jimenez, Luis Emanuel; Alvarez, Rosa Maria Susana; Maffia, Paulo Cesar; Hollmann, Axel; Study of the antibacterial peptide P8.1: Effect on anionic vesicles using spectroscopic techniques; Elsevier Science Inc.; Archives of Biochemistry and Biophysics; 774; 12-2025; 1-410003-9861CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://linkinghub.elsevier.com/retrieve/pii/S0003986125003492info:eu-repo/semantics/altIdentifier/doi/10.1016/j.abb.2025.110635info:eu-repo/semantics/embargoedAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-12-23T13:33:32Zoai:ri.conicet.gov.ar:11336/277472instacron: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:34982025-12-23 13:33:32.554CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Study of the antibacterial peptide P8.1: Effect on anionic vesicles using spectroscopic techniques
title Study of the antibacterial peptide P8.1: Effect on anionic vesicles using spectroscopic techniques
spellingShingle Study of the antibacterial peptide P8.1: Effect on anionic vesicles using spectroscopic techniques
Jimenez, Luis Emanuel
Antimicrobial Peptide
Lipid Phase State Membrane
DLPG
DPPG
title_short Study of the antibacterial peptide P8.1: Effect on anionic vesicles using spectroscopic techniques
title_full Study of the antibacterial peptide P8.1: Effect on anionic vesicles using spectroscopic techniques
title_fullStr Study of the antibacterial peptide P8.1: Effect on anionic vesicles using spectroscopic techniques
title_full_unstemmed Study of the antibacterial peptide P8.1: Effect on anionic vesicles using spectroscopic techniques
title_sort Study of the antibacterial peptide P8.1: Effect on anionic vesicles using spectroscopic techniques
dc.creator.none.fl_str_mv Jimenez, Luis Emanuel
Alvarez, Rosa Maria Susana
Maffia, Paulo Cesar
Hollmann, Axel
author Jimenez, Luis Emanuel
author_facet Jimenez, Luis Emanuel
Alvarez, Rosa Maria Susana
Maffia, Paulo Cesar
Hollmann, Axel
author_role author
author2 Alvarez, Rosa Maria Susana
Maffia, Paulo Cesar
Hollmann, Axel
author2_role author
author
author
dc.subject.none.fl_str_mv Antimicrobial Peptide
Lipid Phase State Membrane
DLPG
DPPG
topic Antimicrobial Peptide
Lipid Phase State Membrane
DLPG
DPPG
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.4
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Antimicrobial peptides (AMPs) are promising complements to antibiotics, yet their membrane-level actions remain incompletely understood. In this work, we characterized how the “de novo” cationic AMP P8.1 interacts with anionic lipid bilayers composed of DPPG (gel phase) or DLPG (fluid phase) using zeta potential, tryptophan and Laurdan fluorescence, Raman microscopy, and a carboxyfluorescein (CF) leakage assay. P8.1 bound both lipids electrostatically, reduced zeta potential, and increased large unilamellar vesicles (LUVs) size. Binding kinetics were faster on DPPG multilamellar vesicles, whereas Trp fluorescence assays showed deeper insertion in DLPG (larger Trp blue-shift and lower acrylamide quenching). Laurdan generalized polarization (GP) increased in DLPG but not in DPPG, indicating reduced water access and higher local order in fluid bilayers. Raman spectra revealed diminished phosphate-band intensity in both systems and, in DLPG, a decreased gauche/trans ratio and narrower 1300 cm-1 band consistent with tighter acyl-chain packing. Difference spectra further showed an amide I shift of P8.1, supporting a random-coil to α-helix transition upon binding to lipids. Finally, P8.1 induced ∼80% CF leakage in DLPG LUVs within minutes. Together, the data indicate that P8.1 engages phosphate groups of lipids and then modulates bilayer structure in a phase- and mechanics-dependent manner—rigidifying short-chain, fluid DLPG and minimally perturbing gel-phase DPPG—providing mechanistic insight relevant to antibacterial activity.
Fil: Jimenez, Luis Emanuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto de Química del Noroeste. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Química del Noroeste; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Química Física; Argentina. Universidad Nacional de Tucumán. Facultad de Ciencias Naturales e Instituto Miguel Lillo; Argentina
Fil: Alvarez, Rosa Maria Susana. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto de Química del Noroeste. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Química del Noroeste; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Química Física; Argentina
Fil: Maffia, Paulo Cesar. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Laboratorio de Biotecnologia y Microbiologia Aplicada ; Secretaria de Investigacion ; Universidad Nacional de Hurlingham;
Fil: Hollmann, Axel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet Noa Sur. Centro de Investigación en Biofísica Aplicada y Alimentos. - Universidad Nacional de Santiago del Estero. Centro de Investigación en Biofísica Aplicada y Alimentos; Argentina
description Antimicrobial peptides (AMPs) are promising complements to antibiotics, yet their membrane-level actions remain incompletely understood. In this work, we characterized how the “de novo” cationic AMP P8.1 interacts with anionic lipid bilayers composed of DPPG (gel phase) or DLPG (fluid phase) using zeta potential, tryptophan and Laurdan fluorescence, Raman microscopy, and a carboxyfluorescein (CF) leakage assay. P8.1 bound both lipids electrostatically, reduced zeta potential, and increased large unilamellar vesicles (LUVs) size. Binding kinetics were faster on DPPG multilamellar vesicles, whereas Trp fluorescence assays showed deeper insertion in DLPG (larger Trp blue-shift and lower acrylamide quenching). Laurdan generalized polarization (GP) increased in DLPG but not in DPPG, indicating reduced water access and higher local order in fluid bilayers. Raman spectra revealed diminished phosphate-band intensity in both systems and, in DLPG, a decreased gauche/trans ratio and narrower 1300 cm-1 band consistent with tighter acyl-chain packing. Difference spectra further showed an amide I shift of P8.1, supporting a random-coil to α-helix transition upon binding to lipids. Finally, P8.1 induced ∼80% CF leakage in DLPG LUVs within minutes. Together, the data indicate that P8.1 engages phosphate groups of lipids and then modulates bilayer structure in a phase- and mechanics-dependent manner—rigidifying short-chain, fluid DLPG and minimally perturbing gel-phase DPPG—providing mechanistic insight relevant to antibacterial activity.
publishDate 2025
dc.date.none.fl_str_mv 2025-12
info:eu-repo/date/embargoEnd/2026-06-06
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/11336/277472
Jimenez, Luis Emanuel; Alvarez, Rosa Maria Susana; Maffia, Paulo Cesar; Hollmann, Axel; Study of the antibacterial peptide P8.1: Effect on anionic vesicles using spectroscopic techniques; Elsevier Science Inc.; Archives of Biochemistry and Biophysics; 774; 12-2025; 1-41
0003-9861
CONICET Digital
CONICET
url http://hdl.handle.net/11336/277472
identifier_str_mv Jimenez, Luis Emanuel; Alvarez, Rosa Maria Susana; Maffia, Paulo Cesar; Hollmann, Axel; Study of the antibacterial peptide P8.1: Effect on anionic vesicles using spectroscopic techniques; Elsevier Science Inc.; Archives of Biochemistry and Biophysics; 774; 12-2025; 1-41
0003-9861
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://linkinghub.elsevier.com/retrieve/pii/S0003986125003492
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.abb.2025.110635
dc.rights.none.fl_str_mv info:eu-repo/semantics/embargoedAccess
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eu_rights_str_mv embargoedAccess
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
dc.publisher.none.fl_str_mv Elsevier Science Inc.
publisher.none.fl_str_mv Elsevier Science Inc.
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
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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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