title: Combination of ellipsometry, laser scanning microscopy and Z-scan fluorescence correlation spectroscopy elucidating interaction of cryptdin-4 with supported phospholipid bilayers.
abstract: The present study has two main objectives. The first is to characterize antimicrobial peptide (AMP) cryptdin-4 (Crp-4) interactions with biological membranes and to compare those interactions with those of magainin 2. The second is to combine the complementary experimental approaches of laser scanning microscopy (LSM), ellipsometry, and Z-scan fluorescence correlation spectroscopy (FCS) to acquire comprehensive information on mechanisms of AMP interactions with supported phospholipid bilayers (SPBs)-a popular model of biological membranes. LSM shows appearance of inhomogeneities in spatial distribution of lipids in the bilayer after treatment with Crp-4. Ellipsometric measurements show that binding of Crp-4 does not significantly change the lipid structure of the bilayer (increase in adsorbed mass without a change in thickness of adsorbed layer). Furthermore, Crp-4 slows the lateral diffusion of lipids within the membrane as shown by Z-scan FCS. All changes of the bilayer induced by Crp-4 can be partially reversed by flushing the sample with excess of buffer. Bilayer interactions of magainin 2 are significantly different, causing large loss of lipids and extensive damage to the bilayer. It seems likely that differences in peptide mode of action, readily distinguished using these combined experimental methods, are related to the distinctive beta-sheet and alpha-helical structures of the respective peptides.
authors.affiliation: J. Heyrovský Institute of Physical Chemistry v.v.i., Academy of Sciences of the Czech Republic, Czech Republic.
authors.forename: Adam
authors.initials: A
authors.lastname: Miszta
authors.forename: Radek
authors.initials: R
authors.lastname: Machán
authors.forename: Ales
authors.initials: A
authors.lastname: Benda
authors.forename: Andre J
authors.initials: AJ
authors.lastname: Ouellette
authors.forename: Wim Th
authors.initials: WT
authors.lastname: Hermens
authors.forename: Martin
authors.initials: M
authors.lastname: Hof
mesh_terms: D008051:Lipid Bilayers / Q000737:chemistry* / Q000378:metabolism; D018613:Microscopy, Confocal / Q000379:methods*; D010743:Phospholipids / Q000737:chemistry* / Q000378:metabolism; D011498:Protein Precursors / Q000737:chemistry / Q000378:metabolism*; D013050:Spectrometry, Fluorescence / Q000379:methods*
chemical_list: D008051:Lipid Bilayers; D010743:Phospholipids; D011498:Protein Precursors; C059569:cryptdin