Macromolecule biosynthesis assay and fluorescence spectroscopy methods to explore antimicrobial peptide mode(s) of action

Publikation: Bidrag til bog/antologi/rapportBidrag til bog/antologiForskningfagfællebedømt

Bimal Jana, Kristin Renee Baker, Luca Guardabassi

Antimicrobial peptides (AMPs) are viable alternatives to the currently available antimicrobials, and numerous studies have investigated their possible use as therapeutic agents for specific clinical applications. AMPs are a diverse class of antimicrobials that often act upon the bacterial cell membrane but may exhibit additional modes of action. Identification of the multiple modes of action requires a comprehensive study at subinhibitory concentrations and careful data analysis since additional modes of action can be eclipsed by AMP action on the cell membrane. Techniques that measure the biosynthesis rate of macromolecules (e.g., DNA, RNA, protein, and cell wall) and the cytoplasmic membrane proton motive force (PMF) energy can help to unravel the diverse modes of action of AMPs. Here, we present an overview of macromolecule biosynthesis rate measurement and fluorescence spectroscopy methods to identify AMP mode(s) of action. Detailed protocols designed to measure inhibition of DNA, RNA, protein, and cell wall synthesis or membrane de-energization are presented and discussed for optimal application of these two techniques as well as to enable accurate interpretation of the experimental findings.

OriginalsprogEngelsk
TitelAntimicrobial Peptides : Methods and Protocols
Antal sider10
Vol/bind1548
ForlagHumana Press
Publikationsdato2017
Sider181-190
ISBN (Trykt)978-1-4939-6735-3
ISBN (Elektronisk)978-1-4939-6737-7
DOI
StatusUdgivet - 2017
NavnMethods in Molecular Biology
Vol/bind1548
ISSN1064-3745

ID: 184357303