Biophysical Analysis to Assess the Interaction of CRAC and CARC Motif Peptides of Alpha Hemolysin of Escherichia coli with Membranes
- Autores
- Cané, Lucía; Guzmán, Fanny; Ballati, Galo; Daza Millone, María Antonieta; Pucci Molineris, Melisa Eliana; Maté, Sabina María; Martini, María Florencia; Herlax, Vanesa Silvana
- Año de publicación
- 2023
- Idioma
- inglés
- Tipo de recurso
- artículo
- Estado
- versión publicada
- Descripción
- Alpha hemolysin of Escherichia coli (HlyA) is a pore-forming protein, which is a prototype of the “Repeat in Toxins” (RTX) family. It was demonstrated that HlyA-cholesterol interaction facilitates the insertion of the toxin into membranes. Putative cholesterol-binding sites, called cholesterol recognition/amino acid consensus (CRAC), and CARC (analogous to CRAC but with the opposite orientation) were identified in the HlyA sequence. In this context, two peptides were synthesized, one derived from a CARC site from the insertion domain of the toxin (residues 341-353) (PEP 1) and the other one from a CRAC site from the domain between the acylated lysines (residues 639-644) (PEP 2), to study their role in the interaction of HlyA with membranes. The interaction of peptides with membranes of different lipid compositions (pure POPC and POPC/Cho of 4:1 and 2:1 molar ratios) was analyzed by surface plasmon resonance and molecular dynamics simulations. Results demonstrate that both peptides interact preferentially with Cho-containing membranes, although PEP 2 presents a lower KD than PEP 1. Molecular dynamics simulation results indicate that the insertion and interaction of PEP 2 with Cho-containing membranes are more prominent than those caused by PEP 1. The hemolytic activity of HlyA in the presence of peptides indicates that PEP 2 was the only one that inhibits HlyA activity, interfering in the binding between the toxin and cholesterol. © 2023 American Chemical Society.
Instituto de Investigaciones Bioquímicas de La Plata - Materia
-
Ciencias Médicas
hemolysin
crac domain
cholesterol - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- http://creativecommons.org/licenses/by-nc-sa/4.0/
- Repositorio
- Institución
- Universidad Nacional de La Plata
- OAI Identificador
- oai:sedici.unlp.edu.ar:10915/162849
Ver los metadatos del registro completo
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Biophysical Analysis to Assess the Interaction of CRAC and CARC Motif Peptides of Alpha Hemolysin of Escherichia coli with MembranesCané, LucíaGuzmán, FannyBallati, GaloDaza Millone, María AntonietaPucci Molineris, Melisa ElianaMaté, Sabina MaríaMartini, María FlorenciaHerlax, Vanesa SilvanaCiencias Médicashemolysincrac domaincholesterolAlpha hemolysin of Escherichia coli (HlyA) is a pore-forming protein, which is a prototype of the “Repeat in Toxins” (RTX) family. It was demonstrated that HlyA-cholesterol interaction facilitates the insertion of the toxin into membranes. Putative cholesterol-binding sites, called cholesterol recognition/amino acid consensus (CRAC), and CARC (analogous to CRAC but with the opposite orientation) were identified in the HlyA sequence. In this context, two peptides were synthesized, one derived from a CARC site from the insertion domain of the toxin (residues 341-353) (PEP 1) and the other one from a CRAC site from the domain between the acylated lysines (residues 639-644) (PEP 2), to study their role in the interaction of HlyA with membranes. The interaction of peptides with membranes of different lipid compositions (pure POPC and POPC/Cho of 4:1 and 2:1 molar ratios) was analyzed by surface plasmon resonance and molecular dynamics simulations. Results demonstrate that both peptides interact preferentially with Cho-containing membranes, although PEP 2 presents a lower KD than PEP 1. Molecular dynamics simulation results indicate that the insertion and interaction of PEP 2 with Cho-containing membranes are more prominent than those caused by PEP 1. The hemolytic activity of HlyA in the presence of peptides indicates that PEP 2 was the only one that inhibits HlyA activity, interfering in the binding between the toxin and cholesterol. © 2023 American Chemical Society.Instituto de Investigaciones Bioquímicas de La Plata2023-06-20info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionArticulohttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://sedici.unlp.edu.ar/handle/10915/162849enginfo:eu-repo/semantics/altIdentifier/issn/0006-2960info:eu-repo/semantics/altIdentifier/doi/10.1021/acs.biochem.3c00164info:eu-repo/semantics/openAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)reponame:SEDICI (UNLP)instname:Universidad Nacional de La Platainstacron:UNLP2025-09-29T11:42:46Zoai:sedici.unlp.edu.ar:10915/162849Institucionalhttp://sedici.unlp.edu.ar/Universidad públicaNo correspondehttp://sedici.unlp.edu.ar/oai/snrdalira@sedici.unlp.edu.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:13292025-09-29 11:42:47.156SEDICI (UNLP) - Universidad Nacional de La Platafalse |
dc.title.none.fl_str_mv |
Biophysical Analysis to Assess the Interaction of CRAC and CARC Motif Peptides of Alpha Hemolysin of Escherichia coli with Membranes |
title |
Biophysical Analysis to Assess the Interaction of CRAC and CARC Motif Peptides of Alpha Hemolysin of Escherichia coli with Membranes |
spellingShingle |
Biophysical Analysis to Assess the Interaction of CRAC and CARC Motif Peptides of Alpha Hemolysin of Escherichia coli with Membranes Cané, Lucía Ciencias Médicas hemolysin crac domain cholesterol |
title_short |
Biophysical Analysis to Assess the Interaction of CRAC and CARC Motif Peptides of Alpha Hemolysin of Escherichia coli with Membranes |
title_full |
Biophysical Analysis to Assess the Interaction of CRAC and CARC Motif Peptides of Alpha Hemolysin of Escherichia coli with Membranes |
title_fullStr |
Biophysical Analysis to Assess the Interaction of CRAC and CARC Motif Peptides of Alpha Hemolysin of Escherichia coli with Membranes |
title_full_unstemmed |
Biophysical Analysis to Assess the Interaction of CRAC and CARC Motif Peptides of Alpha Hemolysin of Escherichia coli with Membranes |
title_sort |
Biophysical Analysis to Assess the Interaction of CRAC and CARC Motif Peptides of Alpha Hemolysin of Escherichia coli with Membranes |
dc.creator.none.fl_str_mv |
Cané, Lucía Guzmán, Fanny Ballati, Galo Daza Millone, María Antonieta Pucci Molineris, Melisa Eliana Maté, Sabina María Martini, María Florencia Herlax, Vanesa Silvana |
author |
Cané, Lucía |
author_facet |
Cané, Lucía Guzmán, Fanny Ballati, Galo Daza Millone, María Antonieta Pucci Molineris, Melisa Eliana Maté, Sabina María Martini, María Florencia Herlax, Vanesa Silvana |
author_role |
author |
author2 |
Guzmán, Fanny Ballati, Galo Daza Millone, María Antonieta Pucci Molineris, Melisa Eliana Maté, Sabina María Martini, María Florencia Herlax, Vanesa Silvana |
author2_role |
author author author author author author author |
dc.subject.none.fl_str_mv |
Ciencias Médicas hemolysin crac domain cholesterol |
topic |
Ciencias Médicas hemolysin crac domain cholesterol |
dc.description.none.fl_txt_mv |
Alpha hemolysin of Escherichia coli (HlyA) is a pore-forming protein, which is a prototype of the “Repeat in Toxins” (RTX) family. It was demonstrated that HlyA-cholesterol interaction facilitates the insertion of the toxin into membranes. Putative cholesterol-binding sites, called cholesterol recognition/amino acid consensus (CRAC), and CARC (analogous to CRAC but with the opposite orientation) were identified in the HlyA sequence. In this context, two peptides were synthesized, one derived from a CARC site from the insertion domain of the toxin (residues 341-353) (PEP 1) and the other one from a CRAC site from the domain between the acylated lysines (residues 639-644) (PEP 2), to study their role in the interaction of HlyA with membranes. The interaction of peptides with membranes of different lipid compositions (pure POPC and POPC/Cho of 4:1 and 2:1 molar ratios) was analyzed by surface plasmon resonance and molecular dynamics simulations. Results demonstrate that both peptides interact preferentially with Cho-containing membranes, although PEP 2 presents a lower KD than PEP 1. Molecular dynamics simulation results indicate that the insertion and interaction of PEP 2 with Cho-containing membranes are more prominent than those caused by PEP 1. The hemolytic activity of HlyA in the presence of peptides indicates that PEP 2 was the only one that inhibits HlyA activity, interfering in the binding between the toxin and cholesterol. © 2023 American Chemical Society. Instituto de Investigaciones Bioquímicas de La Plata |
description |
Alpha hemolysin of Escherichia coli (HlyA) is a pore-forming protein, which is a prototype of the “Repeat in Toxins” (RTX) family. It was demonstrated that HlyA-cholesterol interaction facilitates the insertion of the toxin into membranes. Putative cholesterol-binding sites, called cholesterol recognition/amino acid consensus (CRAC), and CARC (analogous to CRAC but with the opposite orientation) were identified in the HlyA sequence. In this context, two peptides were synthesized, one derived from a CARC site from the insertion domain of the toxin (residues 341-353) (PEP 1) and the other one from a CRAC site from the domain between the acylated lysines (residues 639-644) (PEP 2), to study their role in the interaction of HlyA with membranes. The interaction of peptides with membranes of different lipid compositions (pure POPC and POPC/Cho of 4:1 and 2:1 molar ratios) was analyzed by surface plasmon resonance and molecular dynamics simulations. Results demonstrate that both peptides interact preferentially with Cho-containing membranes, although PEP 2 presents a lower KD than PEP 1. Molecular dynamics simulation results indicate that the insertion and interaction of PEP 2 with Cho-containing membranes are more prominent than those caused by PEP 1. The hemolytic activity of HlyA in the presence of peptides indicates that PEP 2 was the only one that inhibits HlyA activity, interfering in the binding between the toxin and cholesterol. © 2023 American Chemical Society. |
publishDate |
2023 |
dc.date.none.fl_str_mv |
2023-06-20 |
dc.type.none.fl_str_mv |
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http://sedici.unlp.edu.ar/handle/10915/162849 |
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eng |
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eng |
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