Angela Di Somma1, Concetta Avitabile2, Arianna Cirillo3, Antonio Moretta4, Antonello Merlino3, Luigi Paduano3, Angela Duilio5, Alessandra Romanelli6. 1. Department of Chemical Sciences, University of Naples "Federico II" Via Cinthia 4, 80126 Napoli, Italy; National Institute of Biostructures and Biosystems (INBB), Viale Medaglie d'Oro 305, 00136 Roma, Italy. 2. Institute of Biostructures and Bioimaging (CNR), via Mezzocannone 16, 80134 Napoli, Italy. 3. Department of Chemical Sciences, University of Naples "Federico II" Via Cinthia 4, 80126 Napoli, Italy. 4. Department of Sciences, University of Basilicata, Potenza, Italy. 5. Department of Chemical Sciences, University of Naples "Federico II" Via Cinthia 4, 80126 Napoli, Italy. Electronic address: anduilio@unina.it. 6. Department of Pharmaceutical Sciences, University of Milan, Via Venezian 21, 20133 Milan, Italy. Electronic address: alessandra.romanelli@unimi.it.
Abstract
BACKGROUND: The comprehension of the mechanism of action of antimicrobial peptides is fundamental for the design of new antibiotics. Studies performed looking at the interaction of peptides with bacterial cells offer a faithful picture of what really happens in nature. METHODS: In this work we focused on the interaction of the peptide Temporin L with E. coli cells, using a variety of biochemical and biophysical techniques that include: functional proteomics, docking, optical microscopy, TEM, DLS, SANS, fluorescence. RESULTS: We identified bacterial proteins specifically interacting with the peptides that belong to the divisome machinery; our data suggest that the GTPase FtsZ is the specific peptide target. Docking experiments supported the FtsZ-TL interaction; binding and enzymatic assays using recombinant FtsZ confirmed this hypothesis and revealed a competitive inhibition mechanism. Optical microscopy and TEM measurements demonstrated that, upon incubation with the peptide, bacterial cells are unable to divide forming long necklace-like cell filaments. Dynamic light scattering studies and Small Angle Neutron Scattering experiments performed on treated and untreated bacterial cells, indicated a change at the nanoscale level of the bacterial membrane. CONCLUSIONS: The peptide temporin L acts by a non-membrane-lytic mechanism of action, inhibiting the divisome machinery. GENERAL SIGNIFICANCE: Identification of targets of antimicrobial peptides is pivotal to the tailored design of new antimicrobials.
BACKGROUND: The comprehension of the mechanism of action of antimicrobial peptides is fundamental for the design of new antibiotics. Studies performed looking at the interaction of peptides with bacterial cells offer a faithful picture of what really happens in nature. METHODS: In this work we focused on the interaction of the peptide Temporin L with E. coli cells, using a variety of biochemical and biophysical techniques that include: functional proteomics, docking, optical microscopy, TEM, DLS, SANS, fluorescence. RESULTS: We identified bacterial proteins specifically interacting with the peptides that belong to the divisome machinery; our data suggest that the GTPase FtsZ is the specific peptide target. Docking experiments supported the FtsZ-TL interaction; binding and enzymatic assays using recombinant FtsZ confirmed this hypothesis and revealed a competitive inhibition mechanism. Optical microscopy and TEM measurements demonstrated that, upon incubation with the peptide, bacterial cells are unable to divide forming long necklace-like cell filaments. Dynamic light scattering studies and Small Angle Neutron Scattering experiments performed on treated and untreated bacterial cells, indicated a change at the nanoscale level of the bacterial membrane. CONCLUSIONS: The peptide temporin L acts by a non-membrane-lytic mechanism of action, inhibiting the divisome machinery. GENERAL SIGNIFICANCE: Identification of targets of antimicrobial peptides is pivotal to the tailored design of new antimicrobials.
Authors: M D Manniello; A Moretta; R Salvia; C Scieuzo; D Lucchetti; H Vogel; A Sgambato; P Falabella Journal: Cell Mol Life Sci Date: 2021-02-17 Impact factor: 9.261
Authors: Gualtiero Milani; Maria Maddalena Cavalluzzi; Roberta Solidoro; Lara Salvagno; Laura Quintieri; Angela Di Somma; Antonio Rosato; Filomena Corbo; Carlo Franchini; Angela Duilio; Leonardo Caputo; Solomon Habtemariam; Giovanni Lentini Journal: Biomedicines Date: 2021-04-22
Authors: Antonio Moretta; Carmen Scieuzo; Anna Maria Petrone; Rosanna Salvia; Michele Dario Manniello; Antonio Franco; Donatella Lucchetti; Antonio Vassallo; Heiko Vogel; Alessandro Sgambato; Patrizia Falabella Journal: Front Cell Infect Microbiol Date: 2021-06-14 Impact factor: 5.293