title: Noncovalent immobilized artificial membrane chromatography, an improved method for describing peptide-lipid bilayer interactions.
abstract: A promising approach in assessing hydrophobic peptide-membrane interactions is the use of reversed-phase high-performance liquid chromatography. The present study describes the preparation and properties of a noncovalent immobilized artificial membrane (noncovalent IAM) stationary phase. The noncovalent IAM phase was prepared by coating the C18 chains of a reversed-phase HPLC column with the phospholipid ditetradecanoyl-sn-glycero-3-phosphocholine. Lipid coating was achieved by pumping a lipid solution in water-2-propanol through the column. The formation of a bilayer-like structure on the chromatographic surface was confirmed by calculating the phospholipid surface density of the stationary phase. The surface density was determined to be approximately 1.95 mumol m-2, which is close to that of lipid vesicles. The coating was found to be stable in chromatographic elution systems containing less than 35% of acetonitrile. Employing this new technique, we determined interaction parameters of a set of helical antibacterial magainin-2-amide peptides with pairwise substitutions of adjacent amino acids by their D-enatiomers. The results demonstrate that the chromatographic retention behavior of peptides on noncovalent IAM stationary phase shows an excellent correlation with lipid affinities to phospholipid vesicles.
authors.affiliation: Institute of Molecular Pharmacology, Berlin, Germany. ekrause@fmp-berlin.de
authors.forename: E
authors.initials: E
authors.lastname: Krause
authors.forename: M
authors.initials: M
authors.lastname: Dathe
authors.forename: T
authors.initials: T
authors.lastname: Wieprecht
authors.forename: M
authors.initials: M
authors.lastname: Bienert
mesh_terms: D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D008051:Lipid Bilayers / Q000737:chemistry*; D008567:Membranes, Artificial; D010455:Peptides / Q000737:chemistry*; D010743:Phospholipids / Q000737:chemistry
chemical_list: D008051:Lipid Bilayers; D008567:Membranes, Artificial; D010455:Peptides; D010743:Phospholipids