The interaction of antimicrobial peptide anoplin with 1,2-dipalmitoyl-sn-glycero-3-[phospho-rac-(1-glycerol)] lipid monolayers was imaged with atomic force microscopy, scanning transmission X-ray microscopy, and X-ray photoemission electron microscopy. X-ray absorption spectromicroscopy of the surface revealed the domains of the phase-segregated surface to be composed of 98(±5)% lipid while the matrix consisted of a ~50:50 lipid-peptide mixture. We show X-ray spectromicroscopy to be a valuable quantitative tool for label-free imaging of lipid monolayers with antimicrobial peptides at a lateral spatial resolution below 80 nm.
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Anders S, Padmore HA, Duarte RM, Renner T, Stammler T, Scholl A, Scheinfein MR, Stohr J, Seve L, Sinkovic B (1999) Photoemission electron microscope for the study of magnetic materials. Rev Sci Instrum 70:3973–3982
Arseneault M, Bedard S, Boulet-Audet B, Pezolet M (2010) Study of the interaction of lactoferricin b with phospholipid monolayers and bilayers. Langmuir 26:3468–3478
Bechinger B (1999) The structure, dynamics and orientation of antimicrobial peptides in membranes by multidimensional solid-state NMR spectroscopy. Biochim Biophys Acta 1462:157–183
Erbe A, Kerth A, Dathe M, Blume A (2009) Interactions of KLA amphipathic model peptides with lipid monolayers. Chem Bio Chem 10:2884–2892
Fu J, Urquhart SG (2005) Linear dichroism in the X-ray absorption spectra of linear n-alkanes. J Phys Chem A 109:11724–11732
Habermann E (1972) Bees and wasp venom. Science 177:314–322
Hancock REW, Rozek A (2002) Role of membranes in the activities of antimicrobial cationic peptides. FEMS Microbiol Lett 206:143–149
Hancock REW, Sahl HG (2006) Antimicrobial and host-defense peptides as new anti-infective therapeutic strategies. Nature Biotech 24:1551–1557
Hitchcock AP, Dynes JJ, Lawrence JR, Obst M, Swerhone GDW, Korber DR, Leppard GG (2009) Soft X-ray spectromicroscopy of nickel sorption in a natural river biofilm. Geobiology 7:432–453
Kilcoyne ALD, Tylisczak T, Steele WF, Fakra S, Hitchcock P, Franck K, Anderson E, Harteneck B, Rightor EG, Mitchell GE, Hitchcock AP, Yang L, Warwick T, Ade H (2003) Interferometer-controlled scanning transmission X-ray microscopes at the advanced light source. J Synchrotron Rad 10:125–136
Koprinarov IN, Hitchcock AP, McCrory CT, Childs RF (2002) Quantitative mapping of structured polymeric systems using singular value decomposition analysis of soft X-ray images. J Phys Chem B 106:5358–5364
Ladokhin AS, Selsted ME, White SH (1997) Sizing membrane pores in lipid vesicles by leakage of co-encapsulated markers: pore formation by melittin. Biophysical J 72:1762–1766
Leung BO, Hitchcock AP, Brash JL, Scholl A, Doran A, Henklein P, Overhage J, Hilpert K, Hale JD, Hancock REW (2008) An X-ray spectromicroscopy study of competitive adsorption of protein and peptide onto polystyrene-poly(methyl methacrylate). Biointerphases 3:F27–F35
Leung BO, Hitchcock AP, Cornelius R, Brash JL, Doran A, Scholl A (2009a) An X-ray spectromicroscopy study of protein adsorption to a polystyrene-polylactide blend. Biomacromolecules 10:1838–1845
Leung BO, Wang J, Brash JL, Hitchcock AP (2009b) Imaging hydrated albumin on a polystyrene-poly(methyl methacrylate) blend surface with X-ray spectromicroscopy. Langmuir 25:13332–13335
Leung BO, Brash JL, Hitchcock AP (2010) Characterization of biomaterials by soft X-ray spectromicroscopy. Materials 3:3911–3938
Neville F, Ivankin A, Konovalov O, Gildalevitz D (2010) A comparative study on the interactions of SMAP-29 with lipid monolayers. Biochim Biophys Acta 1798:851–860
Nováková E, Mitrea G, Peth C, Thieme J, Mann K, Salditt T (2008) Solid supported multicomponent lipid membranes studied by X-ray spectromicroscopy. Biointerphases 3:FB44–FB54
Park CB, Yi KS, Matsuzaki K, Kim MS, Kim SC (2000) Structure-activity analysis of buforin II, a histone H2A-derived antimicrobial peptide: the proline hinge is responsible for the cell-penetrating ability of buforin II. Proc Natl Acad Sci USA 97:8245–8250
Park SC, Kim JY, Shin SO, Jeong CY, Kim MH, Shin SY, Cheong GW, Park Y, Hahm KS (2006) Investigation of toroidal pore and oligomerization by melittin using transmission electron microscopy. Biochem Biophys Res Comm 343:222–228
Park SC, Kim MH, Hossain MA, Shin SY, Kim Y, Stella L, Wade JD, Park Y, Hahm KS (2008) Amphipathic α-helical peptide, HP (2–20), and its analogues derived from Helicobacter pylori: pore formation mechanism in various lipid compositions. Biochim Biophys Acta 1778:229–241
Peth C, Barkusky F, Sedlmair J, Gleber SC, Novakova E, Neimeyer J, Thieme J, Salditt T, Mann K (2009) Near-edge X-ray absorption fine structure measurements using a laser plasma XUV source. J Phys Conf Ser 186:012032. doi:10.1088/1742-6596/186/1/012032
Shaw JE, Epand RF, Hsu JCY, Mo GCH, Epand RM, Yip CM (2008) Cationic peptide-induced remodelling of model membranes: direct visualization by in situ atomic force microscopy. J Struct Biology 162:121–138
Stewart-Ornstein J, Hitchcock AP, Hernàndez-Cruz D, Henklein P, Overhage J, Hilpert K, Hale J, Hancock REW (2007) Using intrinsic X-ray absorption spectral differences to identify and map peptides and proteins. J Phys Chem B 111:7691–7699
Strang G (1988) Linear algebra and its applications. Harcourt Brace Jovanovich, San Diego
Wang J, Morin C, Li L, Hitchcock AP, Zhang X, Araki T, Doran A, Scholl A (2009) Radiation damage in soft X-ray microscopy. J Electron Spectrosc Rel Phenom 170:25–36
Zhang L, Rozek A, Hancock REW (2001) Interaction of cationic antimicrobial peptides with model membranes. J Biol Chem 276:35714–35722
This research is supported by the Natural Science and Engineering Research Council (NSERC), Early Researcher Award (ERA), Canadian Foundation for Innovation (CFI). X-ray microscopy was carried out using the polymer STXM and magnetic X-PEEM at the ALS, which is supported by the US DoE under contract DE-AC03-76SF00098. We thank David Kilcoyne, Tolek Tyliszczak, and Andrew Doran for their diligence and expertise in keeping the beamlines and microscopes in top condition.
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Leung, B.O., Hitchcock, A.P., Won, A. et al. Imaging interactions of cationic antimicrobial peptides with model lipid monolayers using X-ray spectromicroscopy. Eur Biophys J 40, 805–810 (2011). https://doi.org/10.1007/s00249-011-0690-7
- Antimicrobial peptide
- X-ray microscopy