Protocols for Studying the Interaction of MSI-78 with the Membranes of Whole Gram-Positive and Gram-Negative Bacteria by NMR

  • Nury P. Santisteban
  • Michael R. Morrow
  • Valerie BoothEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 1548)


Antimicrobial peptides (AMPs) may interact with a variety of target cell components, including the lipid bilayer, non-lipidic cell envelope components, and/or intracellular targets. However, most biophysical experiments aimed at elucidating the detailed mechanism of AMPs are limited to simple model membrane systems and neglect potentially functional interactions between AMPs and non-lipidic cell components. One of the biophysical techniques commonly used to study how AMPs interact with lipid bilayers is solid-state deuterium NMR. In this chapter we provide protocols to prepare deuterium-labeled intact Gram-negative and Gram-positive bacteria and to observe these samples using solid-state deuterium NMR. Such experiments have the potential to provide important information about how non-lipidic cell envelope components modulate AMP interactions with the cytoplasmic membrane of bacteria.

Key words

AMP Antimicrobial peptide Solid-state NMR Deuterium NMR Whole-cell NMR 


  1. 1.
    Pius J, Morrow MR, Booth V (2012) 2H solid-state nuclear magnetic resonance investigation of whole Escherichia coli interacting with antimicrobial peptide MSI-78. Biochemistry 51:118–125CrossRefPubMedGoogle Scholar
  2. 2.
    Tardy-Laporte C, Arnold AA, Genard B et al (2013) A H-2 solid-state NMR study of the effect of antimicrobial agents on intact Escherichia coli without mutating. Biochim Biophys Acta 1828:614–622CrossRefPubMedGoogle Scholar
  3. 3.
    Marcotte I, Booth V (2014) 2H solid-state NMR study of peptide-membrane interactions in intact bacteria. In: Advances in biological solid-state NMR: proteins and membrane-active peptides. The Royal Society of Chemistry, Cambridge, pp 459–475CrossRefGoogle Scholar
  4. 4.
    Warnet XL, Laadhari M, Arnold AA et al (2015) A 2H magic-angle spinning solid-state NMR characterisation of lipid membranes in intact bacteria. Biochim Biophys Acta 1858:146–152CrossRefPubMedGoogle Scholar
  5. 5.
    Pius J (2012) A novel approach to characterize the membrane ‐ disrupting activity of antimicrobial peptides using 2 H solid ‐ state NMR of whole Escherichia coli. Master Thesis Memorial University of Newfoundland, St. John’s, NL, CanadaGoogle Scholar
  6. 6.
    Konings AWT, Gipp JJ, Yatvin MB (1984) Radio- and thermosensitivity of E. coli K1060 after thiol depletion by diethylmaleate. Radiat Environ Biophys 23:245–253CrossRefPubMedGoogle Scholar
  7. 7.
    Davis JH, Jeffrey KR, Bloom M et al (1976) Quadrupolar echo deuteron magnetic resonance spectroscopy. In: Ordered hydrocarbon chains. Chem Phys Lett 42:390–394CrossRefGoogle Scholar
  8. 8.
    Prosser RS, Davis JH, Dahlquist FW, Lindorfer MA (1991) Deuterium nuclear magnetic resonance of the gramicidin A backbone in a phospholipid bilayer. Biochemistry 30:4687–4696CrossRefPubMedGoogle Scholar
  9. 9.
    Davis JH (1983) The description of membrane lipid conformation, order and dynamics by 2H-NMR. Biochim Biophys Acta 737:117–171CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  • Nury P. Santisteban
    • 1
  • Michael R. Morrow
    • 1
  • Valerie Booth
    • 1
    • 2
    Email author
  1. 1.Department of Physics and Physical OceanographyMemorial University of Newfoundland and LabradorSt. John’sCanada
  2. 2.Department of BiochemistryMemorial University of Newfoundland and LabradorSt. John’sCanada

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