Noncovalent immobilized artificial membrane chromatography, an improved method for describing peptide–lipid bilayer interactions
| Autor: | Margitta Dathe, Eberhard Krause, Torsten Wieprecht, Michael Bienert |
|---|---|
| Rok vydání: | 1999 |
| Předmět: |
Chromatography
Elution Bilayer Lipid Bilayers Organic Chemistry Synthetic membrane Phospholipid Membranes Artificial General Medicine Biochemistry High-performance liquid chromatography Analytical Chemistry Hydrophobic effect chemistry.chemical_compound chemistry lipids (amino acids peptides and proteins) Peptides Lipid bilayer Chromatography High Pressure Liquid Phospholipids Antibacterial agent |
| Zdroj: | Journal of Chromatography A. 849:125-133 |
| ISSN: | 0021-9673 |
| DOI: | 10.1016/s0021-9673(99)00528-2 |
| Popis: | 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 μmol 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. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect