Peptide:lipid ratio and membrane surface charge determine the mechanism of action of the antimicrobial peptide BP100. Conformational and functional studies

Detalhes bibliográficos
Autor(a) principal: Manzini, Mariana C.
Data de Publicação: 2014
Outros Autores: Perez, Katia Regina [UNIFESP], Riske, Karin Amaral [UNIFESP], Bozelli, Jose C., Santos, Talita L., Silva, Marcia A. da, Saraiva, Greice Kelle Viegas, Politi, Mario J., Valente, Ana P., Almeida, Fabio C. L., Chaimovich, Hernan, Rodrigues, Magali A., Bemquerer, Marcelo P., Schreier, Shirley, Cuccovia, Iolanda M.
Tipo de documento: Artigo
Idioma: eng
Título da fonte: Repositório Institucional da UNIFESP
Texto Completo: http://dx.doi.org/10.1016/j.bbamem.2014.04.004
http://repositorio.unifesp.br/handle/11600/37884
Resumo: The cecropin-melittin hybrid antimicrobial peptide BP100 (H-KKLFKKILKYL-NH2) is selective for Gram-negative bacteria, negatively charged membranes, and weakly hemolytic. We studied BP100 conformational and functional properties upon interaction with large unilamellar vesicles, LUVs, and giant unilamellar vesicles, GUVs, containing variable proportions of phosphatidylcholine (PC) and negatively charged phosphatidylglycerol (PG). CD and NMR spectra showed that upon binding to PG-containing LUVs BP100 acquires a-helical conformation, the helix spanning residues 3-11. Theoretical analyses indicated that the helix is amphipathic and surface-seeking. CD and dynamic light scattering data evinced peptide and/or vesicle aggregation, modulated by peptide: lipid ratio and PG content. BP100 decreased the absolute value of the zeta potential () of LUVs with low PG contents; for higher PG, binding was analyzed as an ion-exchange process. At high salt, BP100-induced LUVS leakage requires higher peptide concentration, indicating that both electrostatic and hydrophobic interactions contribute to peptide binding. While a gradual release took place at low peptide:lipid ratios, instantaneous loss occurred at high ratios, suggesting vesicle disruption. Optical microscopy of GUVs confirmed BP100-promoted disruption of negatively charged membranes. the mechanism of action of BP100 is determined by both peptide:lipid ratio and negatively charged lipid content While gradual release results from membrane perturbation by a small number of peptide molecules giving rise to changes in acyl chain packing, lipid clustering (leading to membrane defects), and/or membrane thinning, membrane disruption results from a sequence of events large-scale peptide and lipid clustering, giving rise to peptide-lipid patches that eventually would leave the membrane in a carpet-like mechanism. (C) 2014 Elsevier B.V. All rights reserved.
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spelling Peptide:lipid ratio and membrane surface charge determine the mechanism of action of the antimicrobial peptide BP100. Conformational and functional studiesBP100Antimicrobial peptideCDNMRZeta potentialModel membrane leakageThe cecropin-melittin hybrid antimicrobial peptide BP100 (H-KKLFKKILKYL-NH2) is selective for Gram-negative bacteria, negatively charged membranes, and weakly hemolytic. We studied BP100 conformational and functional properties upon interaction with large unilamellar vesicles, LUVs, and giant unilamellar vesicles, GUVs, containing variable proportions of phosphatidylcholine (PC) and negatively charged phosphatidylglycerol (PG). CD and NMR spectra showed that upon binding to PG-containing LUVs BP100 acquires a-helical conformation, the helix spanning residues 3-11. Theoretical analyses indicated that the helix is amphipathic and surface-seeking. CD and dynamic light scattering data evinced peptide and/or vesicle aggregation, modulated by peptide: lipid ratio and PG content. BP100 decreased the absolute value of the zeta potential () of LUVs with low PG contents; for higher PG, binding was analyzed as an ion-exchange process. At high salt, BP100-induced LUVS leakage requires higher peptide concentration, indicating that both electrostatic and hydrophobic interactions contribute to peptide binding. While a gradual release took place at low peptide:lipid ratios, instantaneous loss occurred at high ratios, suggesting vesicle disruption. Optical microscopy of GUVs confirmed BP100-promoted disruption of negatively charged membranes. the mechanism of action of BP100 is determined by both peptide:lipid ratio and negatively charged lipid content While gradual release results from membrane perturbation by a small number of peptide molecules giving rise to changes in acyl chain packing, lipid clustering (leading to membrane defects), and/or membrane thinning, membrane disruption results from a sequence of events large-scale peptide and lipid clustering, giving rise to peptide-lipid patches that eventually would leave the membrane in a carpet-like mechanism. (C) 2014 Elsevier B.V. All rights reserved.Univ São Paulo, Inst Chem, Dept Biochem, BR-05513970 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Biophys, BR-04044020 São Paulo, BrazilUniv Fed Rio de Janeiro, Inst Med Biochem, Nucl Magnet Resonance Natl Ctr, Rio de Janeiro, BrazilEmbrapa Recursos Genet & Biotecnol, BR-70770917 Brasilia, DF, BrazilUniversidade Federal de São Paulo, Dept Biophys, BR-04044020 São Paulo, BrazilWeb of ScienceFundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Institut Nacional de Ciencia e Tecnologia de fluidos complexos (INCTFCx)Nude de Apoio Pesquisa de Fluidos Complexos (NAPFCx)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)FAPESP: 2007/50970-5FAPESP: 2013/08166-5Elsevier B.V.Universidade de São Paulo (USP)Universidade Federal de São Paulo (UNIFESP)Universidade Federal do Rio de Janeiro (UFRJ)Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA)Manzini, Mariana C.Perez, Katia Regina [UNIFESP]Riske, Karin Amaral [UNIFESP]Bozelli, Jose C.Santos, Talita L.Silva, Marcia A. daSaraiva, Greice Kelle ViegasPoliti, Mario J.Valente, Ana P.Almeida, Fabio C. L.Chaimovich, HernanRodrigues, Magali A.Bemquerer, Marcelo P.Schreier, ShirleyCuccovia, Iolanda M.2016-01-24T14:37:28Z2016-01-24T14:37:28Z2014-07-01info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersion1985-1999application/pdfhttp://dx.doi.org/10.1016/j.bbamem.2014.04.004Biochimica Et Biophysica Acta-biomembranes. Amsterdam: Elsevier B.V., v. 1838, n. 7, p. 1985-1999, 2014.10.1016/j.bbamem.2014.04.004WOS000336695300034.pdf0005-2736http://repositorio.unifesp.br/handle/11600/37884WOS:000336695300034engBiochimica Et Biophysica Acta-biomembranesinfo:eu-repo/semantics/openAccesshttp://www.elsevier.com/about/open-access/open-access-policies/article-posting-policyreponame:Repositório Institucional da UNIFESPinstname:Universidade Federal de São Paulo (UNIFESP)instacron:UNIFESP2024-10-10T10:58:55Zoai:repositorio.unifesp.br/:11600/37884Repositório InstitucionalPUBhttp://www.repositorio.unifesp.br/oai/requestbiblioteca.csp@unifesp.bropendoar:34652024-10-10T10:58:55Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)false
dc.title.none.fl_str_mv Peptide:lipid ratio and membrane surface charge determine the mechanism of action of the antimicrobial peptide BP100. Conformational and functional studies
title Peptide:lipid ratio and membrane surface charge determine the mechanism of action of the antimicrobial peptide BP100. Conformational and functional studies
spellingShingle Peptide:lipid ratio and membrane surface charge determine the mechanism of action of the antimicrobial peptide BP100. Conformational and functional studies
Manzini, Mariana C.
BP100
Antimicrobial peptide
CD
NMR
Zeta potential
Model membrane leakage
title_short Peptide:lipid ratio and membrane surface charge determine the mechanism of action of the antimicrobial peptide BP100. Conformational and functional studies
title_full Peptide:lipid ratio and membrane surface charge determine the mechanism of action of the antimicrobial peptide BP100. Conformational and functional studies
title_fullStr Peptide:lipid ratio and membrane surface charge determine the mechanism of action of the antimicrobial peptide BP100. Conformational and functional studies
title_full_unstemmed Peptide:lipid ratio and membrane surface charge determine the mechanism of action of the antimicrobial peptide BP100. Conformational and functional studies
title_sort Peptide:lipid ratio and membrane surface charge determine the mechanism of action of the antimicrobial peptide BP100. Conformational and functional studies
author Manzini, Mariana C.
author_facet Manzini, Mariana C.
Perez, Katia Regina [UNIFESP]
Riske, Karin Amaral [UNIFESP]
Bozelli, Jose C.
Santos, Talita L.
Silva, Marcia A. da
Saraiva, Greice Kelle Viegas
Politi, Mario J.
Valente, Ana P.
Almeida, Fabio C. L.
Chaimovich, Hernan
Rodrigues, Magali A.
Bemquerer, Marcelo P.
Schreier, Shirley
Cuccovia, Iolanda M.
author_role author
author2 Perez, Katia Regina [UNIFESP]
Riske, Karin Amaral [UNIFESP]
Bozelli, Jose C.
Santos, Talita L.
Silva, Marcia A. da
Saraiva, Greice Kelle Viegas
Politi, Mario J.
Valente, Ana P.
Almeida, Fabio C. L.
Chaimovich, Hernan
Rodrigues, Magali A.
Bemquerer, Marcelo P.
Schreier, Shirley
Cuccovia, Iolanda M.
author2_role author
author
author
author
author
author
author
author
author
author
author
author
author
author
dc.contributor.none.fl_str_mv Universidade de São Paulo (USP)
Universidade Federal de São Paulo (UNIFESP)
Universidade Federal do Rio de Janeiro (UFRJ)
Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA)
dc.contributor.author.fl_str_mv Manzini, Mariana C.
Perez, Katia Regina [UNIFESP]
Riske, Karin Amaral [UNIFESP]
Bozelli, Jose C.
Santos, Talita L.
Silva, Marcia A. da
Saraiva, Greice Kelle Viegas
Politi, Mario J.
Valente, Ana P.
Almeida, Fabio C. L.
Chaimovich, Hernan
Rodrigues, Magali A.
Bemquerer, Marcelo P.
Schreier, Shirley
Cuccovia, Iolanda M.
dc.subject.por.fl_str_mv BP100
Antimicrobial peptide
CD
NMR
Zeta potential
Model membrane leakage
topic BP100
Antimicrobial peptide
CD
NMR
Zeta potential
Model membrane leakage
description The cecropin-melittin hybrid antimicrobial peptide BP100 (H-KKLFKKILKYL-NH2) is selective for Gram-negative bacteria, negatively charged membranes, and weakly hemolytic. We studied BP100 conformational and functional properties upon interaction with large unilamellar vesicles, LUVs, and giant unilamellar vesicles, GUVs, containing variable proportions of phosphatidylcholine (PC) and negatively charged phosphatidylglycerol (PG). CD and NMR spectra showed that upon binding to PG-containing LUVs BP100 acquires a-helical conformation, the helix spanning residues 3-11. Theoretical analyses indicated that the helix is amphipathic and surface-seeking. CD and dynamic light scattering data evinced peptide and/or vesicle aggregation, modulated by peptide: lipid ratio and PG content. BP100 decreased the absolute value of the zeta potential () of LUVs with low PG contents; for higher PG, binding was analyzed as an ion-exchange process. At high salt, BP100-induced LUVS leakage requires higher peptide concentration, indicating that both electrostatic and hydrophobic interactions contribute to peptide binding. While a gradual release took place at low peptide:lipid ratios, instantaneous loss occurred at high ratios, suggesting vesicle disruption. Optical microscopy of GUVs confirmed BP100-promoted disruption of negatively charged membranes. the mechanism of action of BP100 is determined by both peptide:lipid ratio and negatively charged lipid content While gradual release results from membrane perturbation by a small number of peptide molecules giving rise to changes in acyl chain packing, lipid clustering (leading to membrane defects), and/or membrane thinning, membrane disruption results from a sequence of events large-scale peptide and lipid clustering, giving rise to peptide-lipid patches that eventually would leave the membrane in a carpet-like mechanism. (C) 2014 Elsevier B.V. All rights reserved.
publishDate 2014
dc.date.none.fl_str_mv 2014-07-01
2016-01-24T14:37:28Z
2016-01-24T14:37:28Z
dc.type.driver.fl_str_mv info:eu-repo/semantics/article
dc.type.status.fl_str_mv info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.uri.fl_str_mv http://dx.doi.org/10.1016/j.bbamem.2014.04.004
Biochimica Et Biophysica Acta-biomembranes. Amsterdam: Elsevier B.V., v. 1838, n. 7, p. 1985-1999, 2014.
10.1016/j.bbamem.2014.04.004
WOS000336695300034.pdf
0005-2736
http://repositorio.unifesp.br/handle/11600/37884
WOS:000336695300034
url http://dx.doi.org/10.1016/j.bbamem.2014.04.004
http://repositorio.unifesp.br/handle/11600/37884
identifier_str_mv Biochimica Et Biophysica Acta-biomembranes. Amsterdam: Elsevier B.V., v. 1838, n. 7, p. 1985-1999, 2014.
10.1016/j.bbamem.2014.04.004
WOS000336695300034.pdf
0005-2736
WOS:000336695300034
dc.language.iso.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv Biochimica Et Biophysica Acta-biomembranes
dc.rights.driver.fl_str_mv info:eu-repo/semantics/openAccess
http://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
eu_rights_str_mv openAccess
rights_invalid_str_mv http://www.elsevier.com/about/open-access/open-access-policies/article-posting-policy
dc.format.none.fl_str_mv 1985-1999
application/pdf
dc.publisher.none.fl_str_mv Elsevier B.V.
publisher.none.fl_str_mv Elsevier B.V.
dc.source.none.fl_str_mv reponame:Repositório Institucional da UNIFESP
instname:Universidade Federal de São Paulo (UNIFESP)
instacron:UNIFESP
instname_str Universidade Federal de São Paulo (UNIFESP)
instacron_str UNIFESP
institution UNIFESP
reponame_str Repositório Institucional da UNIFESP
collection Repositório Institucional da UNIFESP
repository.name.fl_str_mv Repositório Institucional da UNIFESP - Universidade Federal de São Paulo (UNIFESP)
repository.mail.fl_str_mv biblioteca.csp@unifesp.br
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