Surface acoustic wave biosensor as a tool to study the interaction of antimicrobial peptides with phospholipid and lipopolysaccharide model membranes.
Surface acoustic wave biosensors are a powerful tool for the study of biomolecular interactions. The modulation of a surface-confined acoustic wave is utilized here for the analysis of surface binding. Phase and amplitude of the wave correspond roughly to mass loading and viscoelastic properties of the surface, respectively. We established a procedure to reconstitute phospholipid and lipopolysaccharide bilayers on the surface of a modified gold sensor chip to study the mode of action of membrane-active peptides. The procedure included the formation of a self-assembled monolayer of 11-mercaptoundecanol, covalent coupling of carboxymethyl-dextran, and subsequent coating with a poly- l-lysine layer. The lipid coverage of the surface is highly reproducible and homogeneous as demonstrated in atomic force micrographs. Ethanol/triton treatment removed the lipids completely, which provided the basis for continuous sequences of independent experiments. The setup was applied to investigate the binding of human cathelicidin-derived peptide LL32, as an example for antimicrobial peptides, to immobilized phosphatidylserine membranes. The peptide-membrane interaction results in a positive phase shift and an increase in amplitude, indicating a mass increase along with a loss in viscosity. This suggests that the bilayer becomes more rigid upon interaction with LL32.
Andrä
Jörg
J
Research Center Borstel, Division of Biophysics, Parkallee 10, Borstel, Germany.
Böhling
Arne
A
Gronewold
Thomas M A
TM
Schlecht
Ulrich
U
Perpeet
Markus
M
Gutsmann
Thomas
T
D000162:Acoustics; D000890:Anti-Infective Agents / Q000737:chemistry*; D015374:Biosensing Techniques / Q000379:methods*; D004926:Escherichia coli / Q000737:chemistry; D008070:Lipopolysaccharides / Q000737:chemistry*; D008567:Membranes, Artificial; D018625:Microscopy, Atomic Force; D008954:Models, Biological; D010455:Peptides / Q000737:chemistry*; D010743:Phospholipids / Q000737:chemistry*; D011513:Proteus mirabilis / Q000737:chemistry; D013499:Surface Properties
D000890:Anti-Infective Agents; D008070:Lipopolysaccharides; D008567:Membranes, Artificial; D010455:Peptides; D010743:Phospholipids