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The antimicrobial peptide Magainin-2 interacts with BamA impairing folding of E. coli membrane proteins

Antimicrobial peptides (AMPs) are a unique and diverse group of molecules endowed with a broad spectrum of antibiotics properties that are being considered as new alternative therapeutic agents. Most of these peptides are membrane-active molecules, killing bacteria by membrane disruption. However, r...

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Autores principales: Di Somma, Angela, Cané, Carolina, Moretta, Antonio, Illiano, Anna, Pinto, Gabriella, Cavasso, Domenico, Amoresano, Angela, Paduano, Luigi, Duilio, Angela
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9620421/
https://www.ncbi.nlm.nih.gov/pubmed/36324521
http://dx.doi.org/10.3389/fchem.2022.1013788
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author Di Somma, Angela
Cané, Carolina
Moretta, Antonio
Illiano, Anna
Pinto, Gabriella
Cavasso, Domenico
Amoresano, Angela
Paduano, Luigi
Duilio, Angela
author_facet Di Somma, Angela
Cané, Carolina
Moretta, Antonio
Illiano, Anna
Pinto, Gabriella
Cavasso, Domenico
Amoresano, Angela
Paduano, Luigi
Duilio, Angela
author_sort Di Somma, Angela
collection PubMed
description Antimicrobial peptides (AMPs) are a unique and diverse group of molecules endowed with a broad spectrum of antibiotics properties that are being considered as new alternative therapeutic agents. Most of these peptides are membrane-active molecules, killing bacteria by membrane disruption. However, recently an increasing number of AMPs was shown to enter bacterial cells and target intracellular processes fundamental for bacterial life. In this paper we investigated the mechanism of action of Maganin-2 (Mag-2), a well-known antimicrobial peptide isolated from the African clawed frog Xenopus laevis, by functional proteomic approaches. Several proteins belonging to E. coli macromolecular membrane complexes were identified as Mag-2 putative interactors. Among these, we focused our attention on BamA a membrane protein belonging to the BAM complex responsible for the folding and insertion of nascent β-barrel Outer Membrane Proteins (OMPs) in the outer membrane. In silico predictions by molecular modelling, in vitro fluorescence binding and Light Scattering experiments carried out using a recombinant form of BamA confirmed the formation of a stable Mag-2/BamA complex and indicated a high affinity of the peptide for BamA. Functional implications of this interactions were investigated by two alternative and complementary approaches. The amount of outer membrane proteins OmpA and OmpF produced in E. coli following Mag-2 incubation were evaluated by both western blot analysis and quantitative tandem mass spectrometry in Multiple Reaction Monitoring scan mode. In both experiments a gradual decrease in outer membrane proteins production with time was observed as a consequence of Mag-2 treatment. These results suggested BamA as a possible good target for the rational design of new antibiotics since this protein is responsible for a crucial biological event of bacterial life and is absent in humans.
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spelling pubmed-96204212022-11-01 The antimicrobial peptide Magainin-2 interacts with BamA impairing folding of E. coli membrane proteins Di Somma, Angela Cané, Carolina Moretta, Antonio Illiano, Anna Pinto, Gabriella Cavasso, Domenico Amoresano, Angela Paduano, Luigi Duilio, Angela Front Chem Chemistry Antimicrobial peptides (AMPs) are a unique and diverse group of molecules endowed with a broad spectrum of antibiotics properties that are being considered as new alternative therapeutic agents. Most of these peptides are membrane-active molecules, killing bacteria by membrane disruption. However, recently an increasing number of AMPs was shown to enter bacterial cells and target intracellular processes fundamental for bacterial life. In this paper we investigated the mechanism of action of Maganin-2 (Mag-2), a well-known antimicrobial peptide isolated from the African clawed frog Xenopus laevis, by functional proteomic approaches. Several proteins belonging to E. coli macromolecular membrane complexes were identified as Mag-2 putative interactors. Among these, we focused our attention on BamA a membrane protein belonging to the BAM complex responsible for the folding and insertion of nascent β-barrel Outer Membrane Proteins (OMPs) in the outer membrane. In silico predictions by molecular modelling, in vitro fluorescence binding and Light Scattering experiments carried out using a recombinant form of BamA confirmed the formation of a stable Mag-2/BamA complex and indicated a high affinity of the peptide for BamA. Functional implications of this interactions were investigated by two alternative and complementary approaches. The amount of outer membrane proteins OmpA and OmpF produced in E. coli following Mag-2 incubation were evaluated by both western blot analysis and quantitative tandem mass spectrometry in Multiple Reaction Monitoring scan mode. In both experiments a gradual decrease in outer membrane proteins production with time was observed as a consequence of Mag-2 treatment. These results suggested BamA as a possible good target for the rational design of new antibiotics since this protein is responsible for a crucial biological event of bacterial life and is absent in humans. Frontiers Media S.A. 2022-10-17 /pmc/articles/PMC9620421/ /pubmed/36324521 http://dx.doi.org/10.3389/fchem.2022.1013788 Text en Copyright © 2022 Di Somma, Cané, Moretta, Illiano, Pinto, Cavasso, Amoresano, Paduano and Duilio. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Chemistry
Di Somma, Angela
Cané, Carolina
Moretta, Antonio
Illiano, Anna
Pinto, Gabriella
Cavasso, Domenico
Amoresano, Angela
Paduano, Luigi
Duilio, Angela
The antimicrobial peptide Magainin-2 interacts with BamA impairing folding of E. coli membrane proteins
title The antimicrobial peptide Magainin-2 interacts with BamA impairing folding of E. coli membrane proteins
title_full The antimicrobial peptide Magainin-2 interacts with BamA impairing folding of E. coli membrane proteins
title_fullStr The antimicrobial peptide Magainin-2 interacts with BamA impairing folding of E. coli membrane proteins
title_full_unstemmed The antimicrobial peptide Magainin-2 interacts with BamA impairing folding of E. coli membrane proteins
title_short The antimicrobial peptide Magainin-2 interacts with BamA impairing folding of E. coli membrane proteins
title_sort antimicrobial peptide magainin-2 interacts with bama impairing folding of e. coli membrane proteins
topic Chemistry
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9620421/
https://www.ncbi.nlm.nih.gov/pubmed/36324521
http://dx.doi.org/10.3389/fchem.2022.1013788
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