Peptide:lipid ratio and membrane surface charge determine the mechanism of action of the antimicrobial peptide BP100. Conformational and functional studies
Autor(a) principal: | |
---|---|
Data de Publicação: | 2014 |
Outros Autores: | , , , , , , , , , , , , , |
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. |
id |
UFSP_d48e71a12158e824afd135c1bdaeb617 |
---|---|
oai_identifier_str |
oai:repositorio.unifesp.br/:11600/37884 |
network_acronym_str |
UFSP |
network_name_str |
Repositório Institucional da UNIFESP |
repository_id_str |
3465 |
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 |
_version_ |
1814268448943898624 |