Biophysical Analysis to Assess the Interaction of CRAC and CARC Motif Peptides of Alpha Hemolysin of Escherichia coli with Membranes

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 recog...

Descripción completa

Guardado en:
Detalles Bibliográficos
Autores principales: Cané, Lucía, Guzmán, Fanny, Ballati, Galo, Daza Millone, María Antonieta, Pucci Molineris, Melisa Eliana, Maté, Sabina María, Martini, María Florencia, Herlax, Vanesa Silvana
Formato: Articulo
Lenguaje:Inglés
Publicado: 2023
Materias:
Ciencias Médicas
hemolysin
crac domain
cholesterol
Acceso en línea:http://sedici.unlp.edu.ar/handle/10915/162849
Aporte de:
SEDICI (UNLP) de Universidad Nacional de La Plata
id I19-R120-10915-162849
record_format dspace
spelling I19-R120-10915-1628492024-02-19T20:09:05Z http://sedici.unlp.edu.ar/handle/10915/162849 Biophysical Analysis to Assess the Interaction of CRAC and CARC Motif Peptides of Alpha Hemolysin of Escherichia coli with Membranes Cané, Lucía Guzmán, Fanny Ballati, Galo Daza Millone, María Antonieta Pucci Molineris, Melisa Eliana Maté, Sabina María Martini, María Florencia Herlax, Vanesa Silvana 2023-06-20 2024-02-19T16:15:24Z en Ciencias Médicas hemolysin crac domain cholesterol 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 Articulo Articulo http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf
institution Universidad Nacional de La Plata
institution_str I-19
repository_str R-120
collection SEDICI (UNLP)
language Inglés
topic Ciencias Médicas
hemolysin
crac domain
cholesterol
spellingShingle Ciencias Médicas
hemolysin
crac domain
cholesterol
Cané, Lucía
Guzmán, Fanny
Ballati, Galo
Daza Millone, María Antonieta
Pucci Molineris, Melisa Eliana
Maté, Sabina María
Martini, María Florencia
Herlax, Vanesa Silvana
Biophysical Analysis to Assess the Interaction of CRAC and CARC Motif Peptides of Alpha Hemolysin of Escherichia coli with Membranes
topic_facet Ciencias Médicas
hemolysin
crac domain
cholesterol
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.
format Articulo
Articulo
author Cané, Lucía
Guzmán, Fanny
Ballati, Galo
Daza Millone, María Antonieta
Pucci Molineris, Melisa Eliana
Maté, Sabina María
Martini, María Florencia
Herlax, Vanesa Silvana
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_sort Cané, Lucía
title Biophysical Analysis to Assess the Interaction of CRAC and CARC Motif Peptides of Alpha Hemolysin of Escherichia coli with Membranes
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
publishDate 2023
url http://sedici.unlp.edu.ar/handle/10915/162849
work_keys_str_mv AT canelucia biophysicalanalysistoassesstheinteractionofcracandcarcmotifpeptidesofalphahemolysinofescherichiacoliwithmembranes
AT guzmanfanny biophysicalanalysistoassesstheinteractionofcracandcarcmotifpeptidesofalphahemolysinofescherichiacoliwithmembranes
AT ballatigalo biophysicalanalysistoassesstheinteractionofcracandcarcmotifpeptidesofalphahemolysinofescherichiacoliwithmembranes
AT dazamillonemariaantonieta biophysicalanalysistoassesstheinteractionofcracandcarcmotifpeptidesofalphahemolysinofescherichiacoliwithmembranes
AT puccimolinerismelisaeliana biophysicalanalysistoassesstheinteractionofcracandcarcmotifpeptidesofalphahemolysinofescherichiacoliwithmembranes
AT matesabinamaria biophysicalanalysistoassesstheinteractionofcracandcarcmotifpeptidesofalphahemolysinofescherichiacoliwithmembranes
AT martinimariaflorencia biophysicalanalysistoassesstheinteractionofcracandcarcmotifpeptidesofalphahemolysinofescherichiacoliwithmembranes
AT herlaxvanesasilvana biophysicalanalysistoassesstheinteractionofcracandcarcmotifpeptidesofalphahemolysinofescherichiacoliwithmembranes
_version_ 1807222464765231104

Ejemplares similares