title: Protocols for Studying the Interaction of MSI-78 with the Membranes of Whole Gram-Positive and Gram-Negative Bacteria by NMR.
abstract: 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.
authors.affiliation: Department of Physics and Physical Oceanography, Memorial University of Newfoundland and Labrador, St. John's, NL, Canada.
authors.forename: Nury P
authors.initials: NP
authors.lastname: Santisteban
authors.affiliation: Department of Physics and Physical Oceanography, Memorial University of Newfoundland and Labrador, St. John's, NL, Canada.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Morrow
authors.affiliation: Department of Physics and Physical Oceanography, Memorial University of Newfoundland and Labrador, St. John's, NL, Canada. vbooth@mun.ca.|Department of Biochemistry, Memorial University of Newfoundland and Labrador, St. John's, NL, Canada, A1B 3X9. vbooth@mun.ca.
authors.forename: Valerie
authors.initials: V
authors.lastname: Booth
mesh_terms: D023181:Antimicrobial Cationic Peptides / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D002462:Cell Membrane / Q000737:chemistry* / Q000378:metabolism; D003903:Deuterium / Q000737:chemistry; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D006090:Gram-Negative Bacteria / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D006094:Gram-Positive Bacteria / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D007553:Isotope Labeling; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D009154:Mutation; D011485:Protein Binding
chemical_list: D023181:Antimicrobial Cationic Peptides; D003903:Deuterium; C093828:pexiganan
keywords: AMP; Antimicrobial peptide; Deuterium NMR; Solid-state NMR; Whole-cell NMR