BioSAXS measurements reveal that two antimicrobial peptides induce similar molecular changes in gram-negative and gram-positive bacteria

Authors Organisations
  • Andreas von Gundlach(Author)
    Ruhr‐Universität Bochum
  • Martin P. Ashby(Author)
    St. George's, University of London
  • Jurnorain Gani(Author)
    St. George's, University of London
  • Paula Matilde Lopez-Perez(Author)
    TiKa Diagnostics Ltd
  • Alan Cookson(Author)
  • Sharon Huws(Author)
  • Christoph Rumancev(Author)
    Ruhr‐Universität Bochum
  • Vasil M. Garamus(Author)
    Helmholtz-Zentrum Geesthacht - Zentrum fűr Material-und Kűstenforschung
  • Ralf Mikut(Author)
    Karlsruhe Institute of Technology
  • Axel Rosenhahn(Author)
    Ruhr‐Universität Bochum
  • Kai Hilpert(Author)
    St. George's, University of London
Type Article
Original languageEnglish
Article number1127
JournalFrontiers in Pharmacology
Volume10
DOI
Publication statusPublished - 26 Sep 2019
Show download statistics
View graph of relations
Citation formats

Abstract

Two highly active short broad-spectrum AMPs (14D and 69D) with unknown mode of action have been investigated in regards to their effect against the Gram-negative bacteria Escherichia coli and the Gram-positive bacteria methicillin-resistant Staphylococcus aureus (MRSA). Minimal inhibitory concentration (MIC) measurements using a cell density of 108 cfu/ml resulted in values between 16 and 32 µg/ml. Time-kill experiments using 108 cfu/ml revealed complete killing, except for 69D in combination with MRSA, where bacterial load was reduced a million times. Small-angle X-ray scattering of biological samples (BioSAXS) at 108 cfu/ml was applied to investigate the ultrastructural changes in E. coli and MRSA in response to these two broad-spectrum AMPs. In addition, electron microscopy (EM) was performed to visualize the treated and non-treated bacteria. As expected, the scattering curves generated using BioSAXS show the ultrastructure of the Gram-positive and Gram-negative bacteria to be very different (BioSAXS is not susceptible to the outer shape). After treatment with either peptide, the scattering curves of E. coli and MRSA cells are much more alike. Whereas in EM, it is notoriously difficult to observe changes for spherical Gram-positives; the BioSAXS results are superior and reveal strongly similar effects for both peptides induced in Gram-positive as well as Gram-negative bacteria. Given the high-throughput possibility and robust statistics, BioSAXS can support and speed up mode of action research in AMPs and other antimicrobial compounds, making a contribution toward the development of urgently needed drugs against resistant bacteria

Keywords

  • antimicrobial peptide, mode of action, SAXS, MRSA, electron microscopy, E. coli